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United States Patent |
5,580,904
|
Ishikura
,   et al.
|
December 3, 1996
|
Formamido and carboxyamido compounds which can be retained in brain
Abstract
According to the present invention, a novel compound group which can pass
through blood-brain barrier (BBB) with carrying a drug thereon and stay
within brain to release the drug and a well-known compound group having
the properties described above are provided.
The compound represented by the general formula
##STR1##
wherein, R.sup.1 represents C.sub.1-6 alkyl which may be substituted by a
group selected from hydroxyl, carboxyl, amino group which may be
substituted by C.sub.1-6 alkyl, and a five- to seven-membered saturated
heterocyclic ring,
R.sup.2 represents hydrogen or C.sub.1-6 alkyl,
R.sup.3 represents hydrogen or C.sub.1-6 alkyl which may be substituted by
hydroxyl,
R.sup.4 represents hydrogen or C.sub.1-6 alkyl,
R.sup.5 represents an amino acid residue, or --S--R.sup.6 or --CO--R.sup.6
wherein R.sup.6 represents C.sub.1-14 alkyl which may be substituted by a
five- to seven-membered saturated ring; C.sub.2-6 alkenyl; aryl; or a
five- to seven-membered saturated ring; or the group represented by the
general formula (IVa):
##STR2##
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 have the same meanings as
defined above, and
.multidot..multidot..multidot..multidot..multidot..multidot..multidot..mult
idot..multidot. represents a single bond or a double bond
provided that at least one of R.sup.1, R.sup.3 and R.sup.5 contains
hydroxyl, carboxyl or amino, and a salt thereof.
Inventors:
|
Ishikura; Toyoaki (Nagareyama, JP);
Ito; Teruomi (Matsudo, JP);
Kato; Takashi (Tsukuba, JP);
Horie; Kazutoshi (Nagareyama, JP);
Ishihara; Hiroshi (Kashiwa, JP);
Senou; Takashi (Adachi-ku, JP)
|
Assignee:
|
Drug Delivery System Institute, Ltd. (Tokyo, JP)
|
Appl. No.:
|
211304 |
Filed:
|
June 24, 1994 |
PCT Filed:
|
July 30, 1993
|
PCT NO:
|
PCT/JP93/01075
|
371 Date:
|
June 24, 1994
|
102(e) Date:
|
June 24, 1994
|
PCT PUB.NO.:
|
WO94/03424 |
PCT PUB. Date:
|
February 17, 1994 |
Foreign Application Priority Data
| Jul 30, 1992[JP] | 4-203897 |
| Jul 30, 1992[JP] | 4-203994 |
Current U.S. Class: |
514/616; 514/629; 564/154; 564/215; 564/224; 568/22 |
Intern'l Class: |
A61K 031/16; C07C 233/01 |
Field of Search: |
514/616,629
564/154,215,224
568/22
|
References Cited
U.S. Patent Documents
5212056 | May., 1993 | Beltramini et al. | 430/572.
|
Foreign Patent Documents |
1-319466 | Dec., 1989 | JP.
| |
2-19365 | Jan., 1990 | JP.
| |
5-134347 | May., 1993 | JP.
| |
Other References
Miyauchi et al., "Studies on orally active cephalosporin esters V. A
prodrug approach for oral delivery of 3-thiazoliomethyl cephalosporin,"
Chemical and Pharmaceutical Bulletin, vol. 38, No. 7, pp. 1906-1910
(1990).
Patent Abstracts of Japan, vol. 11, No. 282 (C-446), Sep. 11, 1987.
Zoltewicz et al., "Preparation and reactivity of model compounds related to
oligomers from thiamin. A mechanism of oligomerisation," Journal of the
American Chemical Society, vol. 103, No. 16, pp. 4900-4904 (1981).
Ostrovskii et al., "Transketolase inhibitors based on disulphide
derivatives of oxythiamin with branched hydrocarbon chains," Chemical
Abstracts, vol. 102, No. 9, abstract No. 77601d (1985).
Zimatkina et al., "Synthesis and study of some new disulphide derivatives
of oxythiamine," Chemical Abstracts, vol. 98, No. 11, abstract No. 89309b
(1983).
Takamizawa et al., "Pyrimidine derivatives and their related compounds,"
Chemical and Pharmaceutical Bulletin, vol., 21, No. 4, pp. 785-790 (1973).
Morita et al., "Active groups in the structure of thiamine
tetrahydrofurfuryl-disulphide in its antipotassium, antiquinidine, and
antiacetylcholine effects on guinea pig atria," Journal of Vitaminology,
vol. 18, No. 4, pp. 225-234 (1972).
|
Primary Examiner: Gupta; Yogendra N.
Attorney, Agent or Firm: Wenderoth, Lind & Ponack
Claims
What is claimed is:
1. A compound represented by the general formula
##STR12##
wherein, R.sup.1 represents C.sub.1-6 alkyl which may be substituted by a
group selected from hydroxyl, carboxyl, and amino group which may be
substituted by C.sub.1-6 alkyl,
R.sup.2 represents hydrogen or C.sub.1-6 alkyl,
R.sup.3 represents hydrogen or C.sub.1-6 alkyl which may be substituted by
hydroxyl,
R.sup.4 represents hydrogen or C.sub.1-6 alkyl,
R.sup.5 represents an amino acid residue, or --S--R.sup.6 or --CO--R.sup.6
wherein R6 represents C.sub.1-14 alkyl which may be substituted by a five-
to seven-membered saturated ring; C.sub.2-6 alkenyl; aryl; C.sub.1-8
alkoxy; or a five- to seven-membered saturated ring, or the group
represented by the general formula (IVa):
##STR13##
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 have the same meanings as
defined above, and
.multidot..multidot..multidot..multidot..multidot..multidot..multidot..mul
t
idot..multidot. represents a single bond or a double bond
provided that at least one of R.sup.1, R.sup.3 and R.sup.5 contains
hydroxyl, carboxyl or amino, and a salt thereof.
2. A compound according to claim 1, wherein
R.sup.1 represents C.sub.1-4 alkyl which may be substituted by a group
selected from hydroxyl, carboxyl, and amino which may be-substituted by a
C.sub.1-4 alkyl group,
R.sup.2 represents hydrogen or C.sub.1-4 alkyl,
R.sup.3 represents hydrogen or C.sub.1-4 alkyl which may be substituted by
hydroxyl,
R.sup.4 represents hydrogen or C.sub.1-4 alkyl,
R.sup.5 represents an alanine residue, a proline residue or --S--R.sup.6 or
--CO--R.sup.6 wherein R.sup.6 represents C.sub.1-14 alkyl which may be
substituted by a five- to seven-membered saturated ring; C.sub.2-6
alkenyl; phenyl; or cyclohexyl; or R.sup.5 represents the group
represented by the formula (IVa) defined above wherein R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 represent the same groups as R.sup.1, R.sup.2, R.sup.3
and R.sup.4 in the formula (Ia)).
3. A method for introducing a drug into the brain of a mammal and retaining
the drugs within the brain, comprising the step of administering a
compound represented by the following formula (IB):
##STR14##
wherein, R.sup.1 represents alkyl or alkenyl, in which one or more
hydrogen atoms in the alkyl group or the alkenyl group may be substituted
by a member selected from the group consisting of hydroxyl which may be
esterified, etherified or carbamated, carboxyl which may be esterified or
amidated, and amino which may be acylated,
A represents the following group:
##STR15##
wherein, R.sup.2 and R.sup.3 represent independently hydrogen, alkyl or
alkenyl, in which one or more hydrogen atoms in the alkyl group or the
alkenyl group may be substituted by a member selected from the group
consisting of hydroxyl which may be esterified, etherified or carbamated,
carboxyl which may be esterified or amidated, and amino which may be
acylated,
m and n are integers of 0 or 1, provided that m and n do not simultaneously
represent 1,
R.sup.11, R.sup.12, R.sup.21 and R.sup.22 independently represent hydrogen,
alkyl or alkenyl, in which one or more hydrogen atoms in the alkyl group
or the alkenyl group may be substituted by a member selected from the
group consisting of hydroxyl which may be esterified, etherified or
carbamated, carboxyl which may be esterified or amidated, and amino which
may be acylated,
R.sup.4 represents hydrogen or alkyl, and
R.sup.5 represents an amino acid residue or a group --X--Y, in which X
represents sulfur or carbonyl, and Y represents alkyl which may be
substituted, alkenyl which may be substituted, or alkoxy which may be
substituted, aryl, a 5- to 7-membered saturated ring, or the following
group (IVA):
##STR16##
wherein R.sup.1, R.sup.4 and A have the same meanings as defined above,
and a salt thereof, wherein the drug is introduced into the compound
through a bond with R.sup.1 to R.sup.5, R.sup.11, R.sup.12, R.sup.21 or
R.sup.22 .
4. The method according to claim 3, wherein
R.sup.1, R.sup.2 and R.sup.3 represent independently alkyl or alkenyl where
the alkyl and alkenyl group may be substituted by a member selected from
the group consisting of hydroxyl; alkylcarbonyloxy which may be
substituted by amino; alkoxy; an amino acid residue; carboxyl;
alkyloxycarbonyl; amino; alkyloxycarbonylamino; phenyl; naphthyl;
cycloalkyl and a five- or six-membered heterocyclic ring which includes up
to 2 hetero atoms selected from oxygen, sulfur and nitrogen and may be
fused together with another ring and may be substituted by C.sub.1-6 alkyl
and/or amino;
provided that the total number of the carbon atoms of R.sup.1, R.sup.2 and
R.sup.3 do not exceed 20,
both m and n are 0, and
Y represents C.sub.1-8 alkyl; or C.sub.2-8 alkenyl; where the alkyl and
alkenyl group may be substituted by a member selected from the group
consisting of amino, alkyloxycarbonylamino, a five- or six-membered
saturated heterocyclic ring containing an oxygen atom or a sulfur atom and
C.sub.5-7 cycloalkyl; or phenyl; naphthyl; or a 5- or 6-membered saturated
heterocyclic ring containing a nitrogen atom.
5. A method for introducing a drug into the brain of a mammal and retaining
the drug within the brain, comprising the step of administering a compound
represented by the following formula (Ib):
##STR17##
wherein, R.sup.1 represents C.sub.1-6 alkyl which may be substituted by a
member selected from the group consisting of hydroxyl, carboxyl, and amino
which may be substituted by C.sub.1-6 alkyl,
R.sup.2 represents hydrogen or C.sub.1-6 alkyl,
R.sup.3 represents hydrogen or C.sub.1-6 alkyl which may be substituted by
hydroxyl,
R.sup.4 represents hydrogen or C.sub.1-6 alkyl,
R.sup.5 represents an amino acid residue, or --S--R.sup.6 or --CO--R.sup.6
wherein R.sup.6 represents C.sub.1-14 alkyl which may be substituted by a
five- to seven-membered saturated ring; C.sub.2-6 alkenyl; aryl; C.sub.1-8
alkoxy; or a five- to seven-membered saturated ring, or the group
represented by the general formula (IVa):
##STR18##
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 have the same meanings as
defined above, and
.multidot..multidot..multidot..multidot..multidot..multidot..multidot..mul
t
idot..multidot. represents a single bond or a double bond
provided that at least one of R.sup.1, R.sup.3 and R.sup.5 contains
hydroxyl, carboxyl, or amino, and a salt thereof, wherein the drug is
introduced into the compound through a bond with R.sup.1, R.sup.2,
R.sup.3, R.sup.4 or R.sup.5.
6. The method according to claim 5, wherein
R.sup.1 represents C.sub.1-4 alkyl which may be substituted by a member
selected from the group consisting of hydroxyl, carboxyl, and amino which
may be substituted by a C.sub.1-4 alkyl group,
R.sup.2 represents hydrogen or C.sub.1-4 alkyl,
R.sup.3 represents hydrogen or C.sub.1-4 alkyl which may be substituted by
hydroxyl,
R.sup.4 represents hydrogen or C.sub.1-4 alkyl,
R.sup.5 represents an alanine residue, a proline residue or --S--R.sup.6 or
--CO--R.sup.6 wherein R.sup.6 represents C.sub.1-14 alkyl which may be
substituted by a five- to seven-membered saturated ring; C.sub.2-6
alkenyl; phenyl; or cyclohexyl; or R.sup.5 represents the group
represented by the formula (IVa) defined above wherein R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 represent the same groups as R.sup.1, R.sup.2, R.sup.3
and R.sup.4 in the formula (Ib).
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a compound which can pass through the
blood-brain barrier (BBB) carrying a drug and release the drug while being
retained in brain, and to its use.
2. Background Art
Cerebral tissue or cells are separated from blood by very tightly combined
cerebral capillary vessels (blood-brain barrier), that is, brain is
protected by the strict limitation of the interchange of materials through
the blood-brain barrier. Therefore, a drug administered systemically
sometimes cannot successfully be delivered to brain being impeded by BBB.
In general, the permeability of materials through BBB is correlated with
the lipophilicity of the materials. Thus, some lipophilic prodrugs have
been synthesized for the purpose of facilitation of delivery to brain of a
drug which is hard to permeate through BBB. On the other hand, brain also
possesses a mechanism for active excretion of waste materials and drugs
back to the periphery. Therefore, it is considered that the conventional
prodrugs which only facilitate their distribution to brain by mainly an
increase in the lipophilicity have limitations in maintaining the
concentration and efficacy of the drug (Rahimy, M. H., et al., Pharm.
Res., 10 (1990) 1061-1067). In other words, it is required to provide a
means to enhance the transport of the drug across BBB and to prolong the
residence in brain of the drug in order to maintain its efficacy following
intravenous administration of a drug.
In order to solve the problem, Bodor (Florida University) has proposed a
dihydropyridine prodrug which utilizes the NAD+-NADH redox system (Bodor,
N., et. al., Science, 214 (1981) 1370-1372). The lipophilic derivative,
when incorporated into cells, is oxidized mainly by the NAD.sup.+
oxidation system into a pyridinium cation and retained in brain due to the
decrease of its permeability through biological membrane. This method is
believed an excellent system for delivering drugs to brain, since the
pyridinium cation carrying the drug releases it in a sustained manner.
Bodor and his co-workers have already demonstrated the excellence of the
method in more than twenty compounds (drugs). However, the dihydropyridine
derivative is easily oxidized per se and thus is likely to lead to the
deterioration of the quality for example by air oxidation after its
synthesis.
The present inventors have examined further possibilities for obtaining the
compounds which can pass through BBB and deliver a drug into brain. As a
result, we have found that certain kinds of compounds have an excellent
property as a carrier for delivering drugs into brain. Thus we have
accomplished the present invention.
SUMMARY OF THE INVENTION
The object of the present invention is to provide novel compounds which can
pass through the blood-brain barrier (BBB) with a drug carried thereon and
release the drug while staying in brain as well as well-known compound
groups which possess the above properties.
According to the present invention, there provides the compound represented
by the formula (IA):
##STR3##
and a salt thereof, wherein
R.sup.1 represents an alkyl group or an alkenyl group, in which one or more
hydrogen atoms in the alkyl group or the alkenyl group may be substituted
by a group selected from the group consisting of a hydroxyl group which
may be esterified, etherified or carbamated, a carboxyl group which may be
esterified or amidated, an amino group which may be acylated and a residue
of cyclic compounds except 4-amino-2-methyl-5-pyridyl group,
A represents the following group:
##STR4##
wherein, R.sup.2 and R.sup.3 represent independently a hydrogen atom, an
alkyl group or an alkenyl group, in which one or more hydrogen atoms in
the alkyl group or the alkenyl group may be substituted by a group
selected from the group consisting of a hydroxyl group which may be
esterified, etherified or carbamated, a carboxyl group which may be
esterified or amidated, an amino group which may be acylated and a residue
of cyclic compounds,
in which R.sup.2 and R.sup.3 in the formula (II) has a cis-configuration,
m and n represent 0 or 1, provided that m and n do not simultaneously
represent 1,
R.sup.11, R.sup.12, R.sup.21 and R.sup.22 independently represent a
hydrogen atom, an alkyl group or an alkenyl group, in which one or more
hydrogen atoms in the alkyl group or the alkenyl group may be substituted
by a group selected from the group consisting of a hydroxyl group which
may be esterified, etherified or carbamated, a carboxyl group which may be
esterified or amidated, an amino group which may be acylated and a residue
of a cyclic compound,
R.sup.4 represents a hydrogen atom or an alkyl group, and
R.sup.5 represents an amino acid residue or a group --X--Y, in which X
represents a sulfur atom or a carbonyl group, and Y represents an alkyl
group which may be substituted or an alkenyl group which may be
substituted or an alkoxy group which may be substituted or a cyclic
compound residue, or the following group (IVA):
##STR5##
wherein R.sup.1, R.sup.4 and A have the same meanings as defined above.
According to the present invention, compounds including some known
compounds represented by the general formula (IB) which can pass through
BBB with a drug supported thereon and release the drug while staying in
brain are also provided in addition to the above novel compounds,
##STR6##
wherein, R.sup.1 represents an alkyl group or an alkenyl group, in which
one or more hydrogen atoms in the alkyl group or the alkenyl group may be
substituted by a group selected from the group consisting of a hydroxyl
group which may be esterified, etherified or carbamated, a carboxyl group
which may be esterified or amidated, an amino group which may be acylated
and a residue of cyclic compounds,
A has the same meaning as defined in the general formula (IA),
R.sup.4 represents a hydrogen atom or an alkyl group, and
R.sup.5 represents an amino acid residue or a group --X--Y, in which X
represents a sulfur atom or a carbonyl group, and Y represents an alkyl
group which may be substituted, an alkenyl group which may be substituted
or an alkoxy group which may be substituted or a cyclic compound residue,
or the following group (IVB):
##STR7##
wherein R.sup.1, R.sup.4 and A have the same meanings as defined above.
DETAILED DESCRIPTION OF THE INVENTION
Novel compounds
In the present specification, alkyl or alkenyl as a group or a part of a
group may be either a straight or branched chain.
In the formula (IA), R.sup.1 represents an alkyl group or an alkenyl group,
and R.sup.2 and R.sup.3 independently represent a hydrogen atom, an alkyl
group or an alkenyl group, in which the total carbon atoms of the alkyl
group or the alkenyl group, which may be appropriately determined in
consideration of the liposolubility of the compound, is preferably not
more than 20. Individual alkyl group or alkenyl group is preferably a
C.sub.1-10 alkyl group or a C.sub.2-10 alkenyl group, more preferably a
C.sub.1-6 alkyl group or a C.sub.2-6 alkenyl group.
One or more hydrogen atoms of the alkyl group and the alkenyl group may be
substituted by a hydroxyl group, a carboxyl group, an amino group or a
cyclic compound residue.
The hydroxyl group as the substituent may also be esterified or etherified.
Furthermore, the carboxyl group may also be esterified or amidated, more
particularly an alkyloxycarbonyl group may be present in place of the
carboxyl group.
The amino group may also be acylated, more particularly an
alkyloxycarbonylamino group may be present in place of the amino group.
The cyclic compound residue for substituting the hydrogen atom of the alkyl
group or the alkenyl group as R.sup.1, R.sup.2 and R.sup.3 includes a
phenyl group, a naphthyl group, a C.sub.5-7 cycloalkyl group and a five-
or six-membered heterocyclic ring which comprises up to 2 hetero atoms
selected from an oxygen atom, a sulfur atom and nitrogen atom, may be
fused together with another ring and may be substituted by a C.sub.1-6
alkyl group and/or an amino group (e.g., a pyridyl group, a pyrimidyl
group, a 4-amino-2-methylpyrimidin-5-yl group, an imidazolyl group, an
indolyl group, a furyl group and a tetrahydrofuryl group), except the case
that the alkyl group in R.sup.1 is replaced by the
4-amino-2-methyl-5-pyrimidyl group.
R.sup.11 R.sup.12, R.sup.21 and R.sup.22 are the groups which are present
when m or n is 1, and specific examples of these groups include preferably
the same ones as R.sup.2 and R.sup.3.
The amino acid residue represented by R.sup.5 preferably bonds as an amino
acid thioester. The amino acid preferably includes alanine, valine,
leucine, isoleucine, phenylalanine, tyrosine, serine, threonine, cysteine,
methionine, aspartic acid, asparagine, glutamic acid, glutamine,
tryptophane and proline.
In the group --X--Y represented by R.sup.5, X represents a sulfur atom or a
carbonyl group, Y represents an alkyl group, preferably a C.sub.1-18 alkyl
group, an alkenyl group, preferably a C.sub.2-8 alkenyl group, an alkoxy
group, preferably C.sub.1-8 alkoxy group, or a cyclic compound residue.
The alkyl group, the alkenyl group and alkoxy group as Y may be substituted
by a substituent. The examples of the substituent include an amino group,
a C.sub.1-7 alkyloxycarbonylamino group, a five- or six-membered saturated
heterocyclic ring containing one of an oxygen atom or a sulfur atom and a
C.sub.5-7 cycloalkyl group.
Moreover, the examples of the cyclic compound residue as Y include a phenyl
group, a naphthyl group, a five- or six-membered heteroaromatic ring
containing a nitrogen atom and a five- or six-membered saturated
heterocyclic ring containing a nitrogen atom.
When R.sup.5 is the group represented by the general formula (IVA), the
compound may be symmetrical at the center of the disulfide bond. R.sup.1
and R.sup.4 present in the right and the left side, respectively, may be
the same or different.
When the group A represents the group (II), the stereochemistry of R.sup.2
and R.sup.3 is of the cis-configuration.
The preferred groups of the compound of the present invention include the
compound in which both m and n are 0.
More preferred groups of the novel compounds according to the present
invention comprise the compound represented by the following formula (Ia):
##STR8##
wherein, R.sup.1 represents a C.sub.1-6 alkyl group which may be
substituted by a group selected from a hydroxyl group, a carboxyl group,
an amino group which may be substituted by a C.sub.1-6 alkyl group, and a
five- to seven-membered saturated heterocyclic ring,
R.sup.2 represents a hydrogen atom or a C.sub.1-6 alkyl group,
R.sup.3 represents a hydrogen atom or a C.sub.1-6 alkyl group which may be
substituted by a hydroxyl group,
R.sup.4 represents a hydrogen atom or a C.sub.1-6 alkyl group,
R.sup.5 represents an amino acid residue, the groups --S--R.sup.6 or
--CO--R.sup.6 in which R.sup.6 represents a C.sub.1-14 alkyl group which
may be substituted by a five- to seven-membered saturated ring; a
C.sub.2-6 alkenyl group; an aryl group; a C.sub.1-8 alkoxy group; or a
five- to seven-membered saturated ring; or the group represented by the
general formula (IVa):
##STR9##
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 have the same meanings as
defined above, and
.multidot..multidot..multidot..multidot..multidot..multidot..multidot..mult
idot..multidot. represents a single bond or a double bond,
with the proviso that at least one of R.sup.1, R.sup.3 and R.sup.5 contains
a hydroxyl group, a carboxyl group or an amino group.
In the formula (Ia), the C.sub.1-6 alkyl as R.sup.1 is preferably a
C.sub.1-4 alkyl group, more preferably a C.sub.1 or C.sub.2 alkyl group.
The alkyl group may be substituted by a hydroxyl group, a carboxyl group,
an amino group which may be substituted by a C.sub.1-6 alkyl group,
preferably a C.sub.1-4 alkyl, more preferably a C.sub.1 or C.sub.2 alkyl;
or a five-to seven-membered saturated heterocyclic ring which preferably
includes a saturated heterocyclic ring comprising a nitrogen atom, more
preferably 1-pyrrolidino, 2-pyrrolidinyl, 3-pyrrolidinyl, piperidino,
2-piperidyl. These substituents are preferably present at the end of an
alkyl group bonded with N.
In the formula (Ia), the C.sub.1-6 alkyl as R.sup.2 is preferably a
C.sub.1-4 alkyl group, more preferably a C.sub.1 or C.sub.2 alkyl group.
In the formula (Ia), the C.sub.1-6 alkyl as R.sup.3 is preferably a
C.sub.1-4 alkyl group, more preferably a C.sub.1 or C.sub.2 alkyl group.
The alkyl group may be substituted by a hydroxyl group, and the preferred
example of the substituted alkyl group is a 2-hydroxyethyl group.
The C.sub.1-6 alkyl as R.sup.4 is preferably a C.sub.1-4 alkyl group, more
preferably a C.sub.1 or C.sub.2 alkyl group.
The amino acid residue as R.sup.5 preferably bonds as an amino acid
thioester. The amino acid preferably includes alanine, valine, leucine,
isoleucine, phenylalanine, tyrosine, serine, threonine, cysteine,
methionine, aspartic acid, asparagine, glutamic acid, glutamine,
tryptophane and proline.
The C.sub.1-14 alkyl group as R.sup.6 in the groups --S--R.sup.6 or
--CO--R.sup.6 represented by R.sup.5 may be either a straight or branched
chain. When the C.sub.1-14 alkyl group is a branched chain, the carbon
atom bonded to --S-- or --CO-- is preferably a secondary or tertiary
carbon atom. The alkyl group may be substituted by a five- to
seven-membered saturated heterocyclic ring which includes preferably a
saturated heterocyclic ring including an oxygen atom or a nitrogen atom,
for example tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidino
and piperidyl. The C.sub.2-6 alkenyl group as R.sup.6 is preferably a
C.sub.2-4 alkenyl group. The aryl group as R.sup.6 is preferably a phenyl
or naphthyl group. The five- to seven-membered saturated ring as R.sup.6
is specifically a cyclopentyl, cyclohexyl or cycloheptyl ring.
When R.sup.6 represents the group represented by the formula (IVa), the
compound can be symmetrical at the center of the disulfide bond (dimer)
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 present in each group may be the
same or different.
In the general formula (Ia) described above,
.multidot..multidot..multidot..multidot..multidot..multidot..multidot..mult
idot..multidot. represents a single bond or a double bond.
In the compound according to the present invention, at least one of the
groups R.sup.1, R.sup.3 and R.sup.5 contains a hydroxyl group, a carboxyl
group or an amino group. When the compound is used as a drug carrier, the
drug is introduced into these functional groups.
Drug Carrier
The compound according to the present invention which can pass through BBB
with a drug carried thereon and release the drug while being retained in
brain is represented by the general formula (IB). Specifically, the
compound group represented by the general formula (IB) comprises the
compounds of the general formula (IA) and compounds of the general formula
(IA) in which R.sup.1 represents an alkyl group substituted by
4-amino-2-methyl-5-pyrimidyl.
A more preferable group of the compound group represented by the general
formula (IB) is a compound represented by the general formula (Ib):
##STR10##
and a salt thereof, wherein,
R*.sup.1 has the same meaning as R1 in claim 1 or represents a C.sub.1-6
alkyl group substituted by a five- or six-membered hetero aromatic ring
which may be substituted by a C.sub.1-4 alkyl or an amino group and
includes one or two nitrogen atoms,
R.sup.2, R.sup.3, R.sup.4 and R.sup.5 have the same meanings as defined in
formula (IB), and at least one of R.sup.3 and R.sup.5 contains a hydroxyl
group, a carboxyl group or an amino group.
As is apparent from the above definitions, the compound group represented
by the general formula (Ib) is the compound group represented by the
general formula (Ia) to which a compound group in which R.sup.1 represents
a C.sub.1-6 alkyl group substituted by a five- or six-membered hetero
aromatic ring containing one or two nitrogen atoms is added. Preferred
examples of the five- or six-membered hetero aromatic ring containing one
or two nitrogen atoms include pyrrolyl, pyridyl, imidazolyl, pyrazolyl,
pyridazinyl, pyrimidyl and pyradinyl. These rings may be substituted by a
C.sub.1-6 alkyl group, preferably a C.sub.1-4 alkyl group, more preferably
a C.sub.1 or C.sub.2 alkyl group and/or an amino group. A preferable
example of R*.sup.1 includes a C.sub.1-4 alkyl group substituted by
4-amino-2-methyl-5-pyrimidyl.
The derivative according to the present invention can be present as a salt
thereof. Examples of the appropriate salts include alkali metal or
alkaline earth metal salts such as a sodium salt, a potassium salt and a
calcium salt, organic ammoniumn salts such as an ammonium salt and a
triethylammonium salt, amino acid salts such as arginine and lysine, a
hydrochloride, a sulfate, a nitrate, a perchlorate, organic sulfonic acid
salts such as a methanesulfonic acid salt, or organic acid salts such as a
succinate, a tartrate and an acetate.
The compound of the formula (I) has a property of passing through BBB with
a drug carried thereon and releasing the drug while being retained in
brain. Without intending to be bound by theory, it is considered that the
compound of the formula (I), after passing through BBB, is cyclized to be
formed as a cation after reduced in the case of the disulfide derivative
or hydrolyzed in the case of the thioester derivative. As a result, the
compound is retained in brain because of decrease in the permeability
through BBB.
Therefore, the compound according to the present invention can be used as a
drug delivery carrier for carrying a drug thereon, permeating through BBB
and delivering the drug into brain.
The drug can be introduced by taking advantage of a variety of functional
groups present in R.sup.1 -R.sup.5 as well as R.sup.11, R.sup.12, R.sup.21
and R.sup.22 of the compound according to the present invention. The drug
is preferably introduced in the form of an acid amide bond, an ester bond
or an urethane bond with no limitation as far as the bond is cleaved and
the drug is released.
The drug which can be introduced is not limited as far as it has a
functional group which can be bonded with a functional group present in
the compound of the formula (IB) in a bonding fashion cleavable in brain.
Also, a drug having no functional groups described above can be modified
by introducing appropriately a functional group so that the drug can form
a bond with the compound of the formula (IB). The embodiment of a drug
which can be introduced includes 3,4-dihydroxy phenylalanine.
It is also said that the lipophilicity and the permeability through BBB of
a low-molecular weight compound are generally correlated with each other.
The compound according to the present invention has also an advantage in
that it has many parts into which a substituent can be introduced and the
lipophilicity or the hydrophilicity of a complex having the drug
introduced thereinto can be further adjusted by appropriately selecting
the substituent.
Furthermore, it can be also said that the compound of the formula (IB) has
a disulfide structure or a thioester structure and is hard to be oxidized.
Synthesis of the drug carrier and introduction of the drug
The compound according to the present invention can be produced by the
following method in accordance with the reports by Matsukawa et al. ((1)
Taizo Matsukawa, Takeo Iwatsu, Hajime Kawasaki, Study of Vitamin B1 and
related compounds (No. 43), Synthesis of Allithiamine Homologues-2,
Yakugaku Zasshi, 73 (1953) 497-501; or (2) Taizo Matsukawa and Hajime
Kawasaki, Study of Vitamin B1 and related compounds (No. 45), Diol type
Vitamin B1 derivatives-1, Yakugaku Zasshi, 73 (1953) 705-708)).
##STR11##
wherein R.sup.1, R.sup.4, A and Y have the same meanings as defined in the
formulae (IA) and (IB), and Z represents a halogen atom such as chlorine,
bromine or iodine or a leaving group such as a tosyl group or a mesyl
group.
The reaction in the step (i) can be effected by mixing the compound of the
formula (V) with R.sup.1 Z in the presence or the absence of of a solvent
such as ethanol or dioxane at room temperature or under heating.
The reaction in the step (ii) is the process for cleaving the cyclic
structure of the formula (VI). The cleavage can be effected by mixing the
compound of the formula (VI) with a sodium hydroxide solution at room
temperature or under heating. The reaction in the subsequent step (iii) is
usually effected without isolating the compound (VII) obtained in this
step to give the compound (VIII).
Among the compounds according to the present invention, the compound (VIII)
of the disulfide type, that is the one of the formula (IB) in which X
represents a sulfur atom, can be obtained via the step (iii). The reaction
can be effected by adding the compound of the formula Y--S--SO.sub.3 Na to
the compound of the formula (VII) with stirring at room temperature. The
compound of the formula Y--S--SO.sub.3 Na can be prepared by heating the
mixture of an aqueous solution of sodium thiosulfate and an ethanolic
solution of the equimolar Y--Z wherein Z represents halogen, under reflux
for 1 to 10 hours.
The completely symmetrical compound of the disulfide type can be prepared
by adding dropwise an aqueous solution of iodine-potassium iodide to the
compound of the formula (VII) with stirring.
Furthermore, the compound of the formula (IB) in which X represents
carbonyl can be prepared by reacting the compound of the formula (VII)
with an acylating agent containing the group Y such as an acid chloride,
an acid anhydride or a sodium acylthiosulfate in an organic solvent such
as ethanol or acetone or in an aqueous solution containing or not
containing inorganic salts such as sodium chloride or sodium sulfate at a
temperature of room temperature to 50.degree. C.
It is needless to say that the functional groups in the aforementioned
synthesis reactions may be protected optionally by an appropriate
protective group.
The drug can be introduced by an appropriate method corresponding to the
functional groups and the functional groups present in the drug. For
instance, in the case of the compound represented by the formula (I) and a
drug containing a carboxyl group, the drug can be introduced by reacting
the drug with the compound represented by the formula (I) under the ester
forming condition. Also, in the case of the compound represented by the
formula (I) and a drug containing an amino group, the drug can be
introduced by reacting these compounds under the carbamate forming
condition. In the case of the compound represented by the formula (I)
containing an amino group and a drug containing a carboxyl group, the drug
can be introduced by reacting these compounds under the amide bond forming
condition. During the reaction of introducing a drug, it is preferred to
protect the other functional groups with appropriate protective groups.
EXAMPLE
The present invention is further illustrated with reference to the
following examples, but it is not limited thereto.
In this connection, the following abbreviations are used in the present
specification:
Compound A: N-dansyl ethylenediamine,
Compound B: N-dansyl .beta.-alanine,
Compound C: N-dansyl L-alanine,
Compound D: N-(tert-butoxycarbonyl)-L-3[3,4-di(pivaloyl-oxy)phenyl]alanine,
and
Boc: tert-butyloxycarbonyl group.
In .sup.1 H-NMR spectrum of the formamide derivative, the compound wherein
R.sup.4 represents a hydrogen atom, the peaks which are probably
attributed to the hydrogens of the N-formyl group are often observed in a
split pattern. Although the ratio of the peaks depends on the compounds,
it varies largely even in the same compound depending on solvents used and
thus presumably is attributed to the rotational isomers of the N-formyl
group. The ratio of the "isomeric mixture" described in the term of 1H-NMR
in examples represents the abundance of the isomers approximately
calculated under the experimental condition.
Intermediate 1:
N-4-Dimethyl-5-[(2-hydroxy)ethyl]thiazolium iodide
Methyl iodide (80 ml) was added to 4-methyl-5-thiazole ethanol (100 g), and
the mixture was heated under reflux for 2 hours. The reaction was
concentrated under reduced pressure to give a dark-brown amorphous
residue. The residue was solidified by adding ether (400 ml) to the
residue. The solid product was further washed twice with ether (500 ml).
The powdery solid products were collected by filtration and dried under
reduced pressure.
Yield: 202 g.
NMR (in D.sub.2 O): .delta.4.10 (3H, s), 3.86 (2H, m), 3.14 (2H, t), 2.48
(3H, s).
Intermediate 2:
N-[(2-hydroxy)ethyl]thiazolium bromide
Thiazole (4.3 g) and 2-bromoethanol (12.5 g) were mixed and heated under
reflux for 5 hours. The reaction was left standing at room temperature to
give white needles. The crystals were washed with acetone, collected by
filtration and dried under reduced pressure.
Yield: 9.53 g.
NMR: (in DMSO-d.sub.6): .delta.10.14 (1H, m), 8.54 & 8.34 (each 1H, dd),
4.62 (2H, t), 3.81 (2H, m).
Intermediate 3:
N-(carboxymethyl)thiazolium bromide
Thiazole (8.5 g), 2-bromoacetic acid (15.0 g) and acetone (20 ml) were
mixed together and left standing at room temperature for 2 days. White
crystals deposited were collected by filtration and washed with acetone.
Yield: 16.96 g.
NMR (in DMSO-d.sub.6): .delta.10.22 (1H, d, J=1.5 Hz), 8.53 & 8.35 (each
1H, each dd), 5.51 (2H, s).
Intermediate 4:
N-[(2-hydroxy)ethyl]-4-methylthiazolium bromide
To the mixture of 4-methylthiazole (24 g) and dioxane (30 ml) was added
2-bromoethanol (40 g), and the resulting mixture was heated under reflux
for 4 hours. To the reaction left standing at room temperature for about
0.5 hour was added acetone (100 ml) with stirring to give white to pale
yellow solids, which were collected by filtration, washed with acetone and
dried under reduced pressure.
Yield: 43.4 g.
NMR (in DMSO-d.sub.6): .delta.10.4 (1H, d), 8.01 (1H, m), 4.53 (2H, t),
3.80 (2H, t), 2.56 (3H,d).
Intermediate 5:
N-[(2-hydroxy)ethyl]-5-methylthiazolium bromide
To the mixture of 5-methylthiazole (10 g) and dioxane (20 ml) was added
2-bromoethanol (15 g), and the resulting mixture was heated under reflux
for 4 hours. To the mixture left standing at room temperature for about
0.5 hour was added acetone (100 ml) with stirring to crystallize pale
yellow solids. The supernatant was discarded, and acetone (50 ml) and
ether (150 ml) were added with stirring to give white to pale yellow
solids, which were collected by filtration, washed with ether and dried
under reduced pressure.
Yield: 19.3 g.
NMR (in DMSO-d.sub.6): .delta.9.96 (1H, d, J=1.5 Hz), 8.32 (1H, t), 4.54
(2H, t), 3.79 (2H, t), 2.57 (3H, d, J=1.5 Hz).
Intermediate 6:
N-[(2-hydroxy)ethyl]-2,4-dimethylthiazolium bromide
To the mixture of 2.4-dimethylthiazole (26.0 g) and dioxane (30 ml) was
added 2-bromoethanol (40 g), and the resulting mixture was heated under
reflux for 5 hours. To the mixture left standing at room temperature for
about 0.5 hour was added acetone (150 ml) with stirring to give pale
yellowish white solids. To the mixture was further added ether (150 ml),
and the resulting powdery crystals were collected by filtration, washed
with acetone and dried under reduced pressure.
Yield: 37.52 g.
NMR (in DMSO-d.sub.6): .delta.7.81 (1H, d), 4.46 (2H, t), 3.79 (2H, t),
3.00 (3H, s), 2.54 (3H, d).
Intermediate 7:
N-5-di-[(2,hydroxy)ethyl]-4-methylthiazolium bromide
To the mixture of 4-methyl-5-thiazole ethanol (29 g) and dioxane (30 ml)
was added 2-bromoethanol (35 g), and the resulting mixture was heated
under reflux for 1.5 hours. To the mixture left standing at room
temperature for about 0.5 hour was added acetone (100 ml) with stirring to
give white to pale yellow solids, which were collected by filtration,
washed with acetone and dried under reduced pressure.
Yield: 42.8 g.
NMR (in DMSO-d.sub.6): .delta.9.92 (1H, s), 4.54 (2H, t), 3.79 (2H, t),
3.65 (2H, t), 3.03 (2H, t).
Intermediate 8:
N-[[2-(tert-butyloxycarbonyl)amino]ethyl]-5-[(2-hydroxy)ethyl]-4-methylthia
zolium bromide
2-Bromoethylammonium bromide (40.0 g) and 4-methyl-5-thiazole ethanol (29.0
g) were dissolved in a hot mixed solution of ethanol (50 ml) and dioxane
(150 ml), and the mixture was heated under reflux for about 30 hours.
Acetone (150 ml) was added to the reaction having been cooled. The mixture
was left standing under ice-cooling for 1 hour to give powdery crystals,
which were collected by filtration, washed with ethanol (150 ml) and dried
under reduced pressure.
Yield: 42.2 g.
NMR (in CD.sub.3 OD): .delta.4.9-4.8 (2H, m), 3.84 (2H, t), 3.59 (2H, t),
3.14 (2H, t), 2.51 (3H, s).
NMR (in DMSO-d.sub.6): .delta.10.04 (1H, s), 4.73 (2H, t), 3.66 (2H, t),
3.40 (2H, broad), 3.04 (2H, t).
The crystals thus obtained (3.5 g) were dissolved in water (50 ml) and
adjusted to a pH of 7-8 with sodium hydrogen carbonate. To this solution
was added with stirring a solution of di-tert-butyl dicarbonate (2.5 g) in
dioxane (50 ml). The stirring was continued at room temperature for 1
hour. Insolubles were removed by filtration, and the filtrate was
concentrated under reduced pressure, diluted with ethanol and concentrated
again. Acetone (20 ml) was added to the residue, and insolubles were
removed by filtration. The filtrate was concentrated under reduced
pressure, and the solution of the residue in a minimum amount of
ethanol/acetone and added dropwise with stirring to a mixed solution (100
ml) of ether: hexane=5:2. The solids crystallized were collected by
filtration and dried under reduced pressure.
Yield: 2.64 g.
NMR (in CD.sub.3 OD): .delta.4.56 (2H, t), 3.81 (2H, t), 3.54 (2H, t), 3.11
(2H, t), 2.57 (3H, s), 1.36 (9H, s).
NMR (in DMSO-d.sub.6): .delta.9.89 (1H, s), 4.50 (2H, t), 3.63 (2H, dd),
3.40 (2H, dd), 3.01 (2H, t), 2.47 (3H, s), 1.31 (9H, s).
Intermediate 9:
N-[2-(Dimethylamino)ethyl]-5-[(2-hydroxy)ethyl]-.sub.4 -methylthiazolium
chloride hydrochloride
To a mixture of 4-methyl-5-thiazole ethanol (16.0 g) and dimethylaminoethyl
chloride hydrochloride (14.4 g) was added ethanol (10 ml. The mixture was
heated under reflux for about 20 hours. Ethanol (10 ml) and acetone (100
ml) were added, and the mixture was stirred to give pale yellow powdery
crystals, which were collected by filtration, washed with acetone
containing a small amount of ethanol and dried under reduced pressure.
Yield: 17.1 g.
NMR (in D.sub.2 O): .delta.4.94 (2H, t), 3.86 (2H, broad t), 3.72 (2H, t),
3.16 (3H, broad t), 3.02 (6H, s), 2.56 (3H, s).
Intermediate 10:
N-[2-(1-pyrrolidino)ethyl]-5-[(2-hydroxy)ethyl-4-methylthiazolium chloride
hydrochloride
To a mixture of 4-methyl-5-thiazole ethanol (14.3 g) and
2-(1-pyrrolidino)ethyl chloride hydrochloride (13.6 g) was added ethanol
(10 ml). The mixture was heated under reflux for about 20 hours. After
cooling, the mixture was added with ethanol (10 ml) and acetone (100 ml),
and stirred to give pale yellow powdery crystals, which were collected by
filtration, washed with acetone containing a small amount of ethanol and
dried under reduced pressure.
Yield: 16.2 g.
NMR (in DMSO-d.sub.6): .delta.10.30 (1H, s), 4.98 (2H, t), 3.71 (2H, m),
3.64 (2H, t), 3.57 (2H, m), 3.08 (2H, m), 3.03 (2H, t), 2.54 (3H, s), 2.02
& 1.89 (each 2H, each m).
Intermediate 11:
N-(2-hydroxypropyl)-4-methylthiazolium chloride hydrochloride
To a mixture of 4-methylthiazole (10.0 g) and dioxane (20ml) was further
added 1-bromo-2-propanol (containing 20% of 2-bromo-1-propanol) (15.3 g),
and the mixture was heated under reflux for about 6 hours. After cooling,
the reaction was added with acetone (30 ml) and ether (130 ml) and left
standing. After the pale brown supernatant was discarded, ether (150 ml)
was further added to the amorphous precipitate to solidify it. Pale brown
powder thus crystallized was collected by filtration and dried under
reduced pressure.
Yield: 8.74 g.
NMR (in DMSO-d.sub.6): .delta.10.04 (1H, d, J=3 Hz), 8.01 (1H, q), 4.52
(1H, dd), 4.28 (1H, dd), 4.00 (1H, m), 2.56 (3H, d, J=1 Hz), 1.19 (3H, d,
J=6 Hz).
Intermediate 12:
N-(2-hydroxyethyl)-2-methylthiazolium bromide
To a solution of 2-methylthiazoline (10.1 g) in dioxane (20 ml) was added
2-bromoethanol (13.8 g), and the mixture was heated under reflux for about
6 hours. After cooling the reaction, acetone (30 ml) and ether (130 ml)
were added with stirring, and the resulting mixture was left standing.
After the pale brown supernatant was discarded, ether (150 ml) was further
added to the amorphous precipitate thus obtained. The mixture was stirred
and left standing. Supernatant obtained was discarded, and the viscous
precipitates obtained was left standing at an ambient temperature under
reduced pressure to give the title compound as the yellow amorphous
residue.
Yield: 19.5 g.
NMR (in DMSO-d.sub.6): .delta.4.51 (2H, t), 3.87 (2H, t), 3.72 (2H, t),
3.67 (2H, t), 2.61 (3H, s).
Example 1
N-methyl-N-[4-hydroxy-1-methyl-2-[(ethyl)dithio]-1-butenyl]formamide
To a solution of sodium hydroxide (8.0 g) in distilled water (50 ml) was
added Intermediate 1 (28.6 g), and the mixture was left standing at room
temperature for 10 minutes, during which sodium sulfate was added to
saturate the reaction solution. To this aqueous solution was added sodium
ethylthiosulfate (46 g) in the form of powder. After the mixture was
stirred at room temperature for 10 minutes, it was extracted with ethyl
acetate (100 ml). The ethyl acetate layer was dried over anhydrous sodium
sulfate and concentrated under reduced pressure. The residue thus obtained
was purified by silica gel column chromatography to give a colorless oil
product (10.3 g).
1H-NMR (in CDCl.sub.3): .delta.7.98 & 7.93 (1H, s), 3.82 (2H, m), 2.96 (3H,
s), 2.88 (2H, t), 2.63 (2H, q), 2.01 (3H, s), 1.28 (3H, s).
Example 2
(a) N-methyl-N-[4-hydroxy-1-methyl-2-[(1-propyl)dithio]-1-butenyl]formamide
To a solution of sodium hydroxide (2.0 g) in distilled water (50 ml) was
added Intermediate 1 (7.2 g), and the mixture was left standing for 10
minutes. To this aqueous solution was added sodium 1-propylthiosulfate (8
g) in the form of powder. A pale yellow oil product was deposited
immediately after the addition of sodium 1-propylthiosulfate. The reaction
mixture was extracted with ethyl acetate (200 ml). The ethyl acetate layer
was dried over anhydrous sodium sulfate and concentrated under reduced
pressure. The residue thus obtained was purified by silica gel column
chromatography to give a colorless oil product (3.9 g).
1H-NMR (in CD.sub.30 D ): .delta.7.98 & 7.90 (1H, s), 3.72 (2H, t), 2.94
(3H, s), 2.87 (2H, t), 2.62 (2H, t), 2.03 (3H, s), 1.67 (2H, m), 0.98 (3H,
t).
(b)
N-methyl-N-[4-[2-[(5-dimethylaminonaphtylsulfonyl)amino]ethylaminocarbonyl
oxy]-1-methyl-2-[(1-propyl)dithio]-1-butenyl]formamide
To a solution of the compound in the above step (a) (3.86 g) in
tetrahydrofuran (70 ml) was added carbonyldiimidazole (2.6 g) in the form
of powder to form a solution in a water bath at 40.degree. C. (for about
10 minutes). To this solution were added Compound A (5.0 g), and the
mixture was stirred at room temperature for 15 minutes. After insolubles
was removed by filtration, the filtrate was evaporated under reduced
pressure. The residue thus obtained was purified by silica gel column
chromatography (CHC13:MeOH=80:1.fwdarw.50:1). To a solution of the
fluorescent yellowish green solid residue (5.34 g) in ethanol (200 ml) was
added 1N HCl (9.4 ml), and the mixture was evaporated under reduced
pressure. Evaporation was repeated further twice by adding ethanol to the
residue. A solution of the residue thus obtained in an ether containing a
small amount of ethanol was added dropwise to ether (300 ml), and the
fluorescent yellow solids crystallized was collected by filtration and
dried under reduced pressure.
Yield: 3.7 g.
1H-NMR (in CD.sub.3 OD): .delta.8.76, 8.49 & 8.34 (each 1H, each d), 7.89
(1H, s), 7.85-7.79 (2H, m), 4.17 (2H, t), 3.35 (6H, s), 3.08-3.05 (3H, m),
2.98-2.91 (3H, m), 2.90 (3H, s), 2.61 (2H, t), 1.99 (3H, s), 1.65 (2H, m),
0.97 (3H, t).
Example 3
(a) N-methyl-N-[4-hydroxy-1-methyl-2-[(2-propyl)dithio]-1-butenyl]formamide
To a solution of sodium hydroxide (8.0 g) in water (50 ml) was added
Intermediate 1 (28.6 g), and the mixture was left standing at room
temperature for 10 minutes. To this aqueous solution was added sodium
2-propylthiosulfate (43 g) in the form of powder. A pale yellow oil
product was deposited immediately after the addition of sodium
2-propylthiosulfate. After the reaction mixture was extracted with ethyl
acetate (200 ml), the ethyl acetate layer was dried over anhydrous sodium
sulfate and concentrated under reduced pressure.
Yield: 24.9 g.
1H-NMR (in CDCl.sub.3): .delta.7.99 & 7.95 (0.33 & 0.67H, each s), 3.78
(2H, t), 2.97 (3H, s), 2.93-2.87 (3H, m), 2.00 (3H, s), 1.27 (6H, d).
(b)
N-methyl-N-[[4-[2-amino-3-(3,4-dipivaloyloxyphenyl)propionyloxy]-1-methyl-
2-[(2-propyl)dithio]-1-butenyl]formamide
The compound in the above step (a) (3.72 g), Compound D (7.0 g) and
dimethylaminopyridine (245 mg) were dissolved in acetonitrile (30 ml). The
mixed solution was ice-cooled. Dicyclohexylcarbodiimide (3.2 g) was added
to the mixed solution and left standing for 3 hours with ice-cooling. In
TLC (CHCl.sub.3 :MeOH=10:1), the spot of the compound in the
above-described step (a) disappeared and converged on the spot at the Rf
value of 0.7. After deposited products were removed by filtration, the
filtrate was concentrated under reduced pressure, and the residue was
purified by silica gel column chromatography (CHC.sub.3
:MeOH=50:1.fwdarw.40:1) to give the protected product of the title
compound.
Yield: 8.26 g.
1H-NMR (in CDCl.sub.3): .delta.7.90 (1H, s), 7.06-6.91 (3H, m), 5.01 (1H,
broad d), 4.54 (1H, broad m), 4.27 & 4.15 (each 1H, each m), 3.06 (2H, d),
2.94 (3H, s), 2.92-2.86 (3H, m), 1.93 (3H, s), 1.43 (9H, s), 1.330 & 1.328
(each 9H, each s), 1.26 (6H, d).
The protected product (4.60 g) was dissolved in trifluoroacetic acid under
ice-cooling. After left standing at room temperature for 15 minutes, the
solution was added dropwise to a sodium hydrogen carbonate suspension (200
ml) to give a white amorphous product. The mixture was extracted with
chloroform. The chloroform layer was dried over anhydrous sodium sulfate
and concentrated under reduced pressure. After the colorless residue thus
obtained was dissolved in ether, n-hexane was added to the ethereal
solution. The mixture was concentrated under reduced pressure in a
low-temperature water bath to give white powder. The powder was collected
by filtration and dried under reduced pressure to give the title compound.
Yield: 3.77 g.
1H-NMR (in CDCl.sub.3): .delta.7.91 (1H, s), 7.06 (2H, s), 6.98 (1H, s),
4.28-4.18 (2H, m), 3.70 (1H, q), 3.10-3.02 (1H, m), 2.95 (3H, s),
2.94-2.82 (4H, m), 1.96 (3H, s), 1.334 & 1,332 (each 9H, each s), 1.26
(6H, d).
(c)
N-methyl-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylaminocarbony
loxy]-1-methyl-2-[(2-propyl)dithio]-1-butenyl]formamide
To the solution of compound in the above step (a) (2.5 g) in
tetrahydrofuran (15 ml) was added carbodiimidazole (1.78 g, 11 mmole) in
the form of powder, and the carbodiimidazole was dissolved in a water bath
at 40.degree. C. (5 minutes). To this solution were added Compound A (2.9
g) and tetrahydrofuran (30 ml), and the mixture was stirred at room
temperature for about 2 hours. After insolubles were removed by
filtration, the filtrate was concentrated under reduced pressure and
purified by silica gel column chromatography (CHCl.sub.3 :MeOH=40:1). To a
solution of the fluorescent yellowish green residue thus obtained (4.95 g)
in ethanol (100 ml) was added 1N hydrochloric acid (8.7 ml), and the
mixture was evaporated under reduced pressure. Dissolution in ethanol and
evaporation was further repeated twice. Ether was added to the residue
obtained, and the resulting white powdery solid was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 4.63 g.
1H-NMR (in DMSO-d.sub.6): .delta.8.53 & 8.37 (each 1H, each d), 8.12 (1H,
m), 7.79 (1H, s), 7.68 & 7.66 (each 1H, each t), 7.44 (1H, d), 4.03 (2H,
t), 2.99-2.90 (9H, m), 2.82 (3H, s), 2.82-2.78 (4H, m), 1.90 (3H, s), 1.19
(6H, d).
Example 4
N-Methyl-N-[4-hydroxy-1-methyl-2-[(2-butyl)dithio]-1-butenyl]formamide
To a solution of sodium hydroxide (3.2 g) in distilled water (10 ml) was
added an aqueous solution of Intermediate 1 (11.4 g), and the mixture was
left standing at room temperature for 30 minutes. Sodium
2-butylthiosulfate (15 g) in the form of powder was added to the aqueous
solution. Immediately after the addition, a light brown oil product was
deposited. The reaction mixture was extracted with ethyl acetate (200 ml).
The ethyl acetate layer was dried over anhydrous sodium sulfate and
concentrated under reduced pressure. The residue thus obtained was
purified by silica gel column chromatography to give a colorless oil
product (6.2 g).
.sup.1 H-NMR (in CD.sub.3 OD): 7.98 & 7.92 (1H, s), 3.71 (2H, t), 2.95 (3H,
s), 2.88 (2H, t), 2.70 (1H, q), 2.02 (3H, s), 1.66 (1H, m), 1.51 (1H, m),
1.25 (3H, d), 0.97 (3H, d).
Example 5
(a) N-(2-hydrxyethyl)-N-[-2-[(1-methylbutyl)dithio]vinyl]-formamide
In a solution of sodium hydroxide (1.32 g) in water (50 ml) was dissolved
Intermediate 2 (3.2 g). To this solution was added sodium
1-methylbutylthiosulfate (6 g) in the form of powder with stirring. After
the mixture turned white turbid, a pale yellow oil product was deposited
and sedimented. The reaction was extracted with chloroform. The chloroform
layer was dried over anhydrous sodium sulfate. The solvent was evaporated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography (CHCl.sub.3 :MeOH=40:1.fwdarw.30:1).
Yield: 2.33 g.
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.33 & 8.09
(0.75 & 0.25H, each s), 6.47, 6.13, 5.91 & 5.82 (0.25, 0.75, 0.75 & 0.25H,
each d, J=9, 8, 8, 9 Hz), 3.82-3.74 (4H, m), 2.92 (1H, m), 2.14 (1H,
broad), 1.63(1H, m), 1.45 (3H, m), 1.33 & 1.32 (3H, each d, each J=7.0
Hz), 0.92 (3H, t).
(b)
N-[2-[3-[(5-dimethylaminonaphthylsulfonyl)amino]-propionyloxy]ethyl]-N-[2-
[(s-butyl)dithio]vinyl]formamide
The compound in the above step (a) (2.31 g), Compound B (3.10 g) and
dimethylaminopyridine (250 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (2.02 g) was added to the solution. The mixture
was then left standing for 15 hours at room temperature. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :MeOH=10:1.fwdarw.8:1).
Yield: 4.77 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.54 & 8.30
(each 1H, each d), 8.26 & 7.91 (0.8 & 0.2H, each s), 8.25 (1H, dd),
7.60-7.51 (2H, m), 7.19 (1H, d), 6. 43, 6.02, 5.87 & 5.77 (0.2, 0.8, 0.8 &
0.2H, each d, J=9, 8, 8, 9 Hz), 5.60 & 5.31 (0.8 & 0.2H, broad t), 4.15
(2H, t), 3.79 (2H, t), 3.17 (2H, q), 2.90 (7 H, m), 2.45 (2H, t), 1.62
(1H, m), 1.44 (3H, m), 1. 31 & 1.30 (3H, each d, each J=7 Hz), 0.91 (3H,
t).
Example 6
(a) N-(carboxymethyl)-N-[2-[(1-methylbutyl)dithio]vinyl]formamide
In a solution of sodium hydroxide (1.32 g) in water (40 ml) was dissolved
Intermediate 3 (2.3 g), and the resulting solution was further saturated
with sodium sulfate. To this solution was added sodium
1-methylbutylthiosulfate (6 g) in the form of powder. The mixture was then
stirred at room temperature for 15 minutes. The reaction was adjusted to a
pH of about 2 with 4N hydrochloric acid and extracted with ethyl acetate
(150 ml). After the ethyl acetate layer was dried over anhydrous sodium
sulfate, and the solvent was evaporated under reduced pressure to give
pale yellow crystals.
Yield: 1.80 g.
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.40 & 8.37
(0.25 & 0.75H, each s), 6.96, 6.22, 5.86 & 5.69 (0.25, 0.75, 0.75 & 0.25H,
each d, J=13.5, 8.5, 8.5, 13.5 Hz), 4.49 & 4.39 (1.5 & 0.5H, each s), 2.91
(1H, m), 1.63(1H, m), 1.44 (3H, m), 1.32 (3H, d), 0.92 (3H, t).
(b)
N-[[[12-[(5-dimethylaminonaphthylsulfonyl)amino]ethyl]-aminocarbonyl]methy
l]-N-[2-[(1-methylbutyl)dithio]vinyl]formamide
Compound A (1.7 g), the compound in the above step (a) (1.5 g) and
dimethylaminopyridine (120 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (1.23 g) was added to the solution. The mixture
was then stirred at room temperature for about 15 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.10:2).
Yield: 1.10 g.
.sup.1 H-NMR (in CDCl.sub.3, 2:1 isomeric mixture): .delta.8.56 (1H, m),
8.40 & 8.31 (0.33 & 0.67H, each s), 8.27-8.21 (2H, m), 7.61-7.51 (2H, m),
7.20 (1H, d), 6.46 & 6.37 (0.33 & 0.67H, each broad), 6.96, 6.08, 5.89 &
5.82 (0.33, 0.67, 0.67 & 0.33H, each d, J=13.5, 8.0, 8.0 & 13.5 Hz), 5.40
& 5.30 (0.67 & 0.33H, each broad), 4.17 & 4.11 (0.67 & 1.33H), 3.32 & 3.04
(each 2H, each m), 2.93 (1H, m), 2.90 (6H, s), 1.61 (1H, m), 1.43 (3H, m),
1.311 & 1.306 (3H, each d, each J=7 Hz), 0.91 (3H, m).
Example 7
(a) N-(2-hydroxyethyl)-N-[1-methyl-2-(2-propyldithio)vinyl]formamide In a
solution of sodium hydroxide (1.32 g) in water (40 ml) was dissolved
Intermediate 4 (3.4 g). To this solution was added sodium
2-propylthiosulfate (ca. 7 g) in the form of powder, and the mixture was
stirred at room temperature for 15 minutes. The reaction was extracted
with chloroform. After the chloroform layer was dried with anhydrous
sodium sulfate, the solvent was evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :MeOH=40:1.fwdarw.35:1).
Yield: 2.41 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): 8.10 & 8.08 (0.8 &
0.2H, each s), 6.20 & 6.07 (0.2 & 0.8H, each d, each J=1.0 Hz), 3.79 &
3.68 (1.6 & 0.4H, each m), 3.65 & 3.54 (1.6 & 0.4H, each t), 06 (1H, m),
2.54 & 2.22 (0.7 & 0.1H, each t), 1.96 & 1.93 (2.4 & 0.6H, each d, each
J=1.0 Hz), 1.33 & 1.32 (6H, each d, each J=7.0 Hz).
.sup.1 H-NMR (in DMSO-d.sub.6, 7:3 isomeric mixture): 8.1 & 7.97 (0.3 &
0.7H, each s), 6.13 & 6.10 (0.7 & 0.3H, each d, each J=1.0 Hz), 4.78 &
4.73 (0.3 & 0.7H, each broad t), 3.48-3.42 (4H, m), 3.10 (1H, m), 1.96 &
1.89 (2.1 & 0.9H, each d, J=1.0 Hz), 1.25 & 1.24 (6H, each d, each J=6.5
Hz).
(b)
N-[2-[2-[(5-dimethylaminonaphthylsulfonyl)amino]propionyloxy]ethyl-N-[1-me
thyl-2-(2-propyldithio)vinyl]formamide
Compound C (2.58 g), the compound in the above step (a) (1.90 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (1.70 g) was added to the solution. The mixture
was then stirred at room temperature for about 12 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1).
Yield: 3.51 g.
.sup.1 H-NMR (in CDCl.sub.3, 7:1 isomeric mixture): .delta.8.54 & 8.30
(each 1H, each d), 8.24 (1H, dd), 8.03 & 7.87 (0.87 & 0.13H, each s),
7.61-7.50 (2H, m), 7.19 (1H, d), 6.10 & 6.02 (0.13 & 0.87H, each d, each
J=1.0 Hz), 5.46 & 5.34 (0.87 & 0.13H, each d, each J=8.5 Hz), 3.98 (1H,
m), 3.93 & 3.87 (each 1H, each m), 3.51 (2H, m), 3.04 (1H, m), 2.88 (6H,
s), 1.85 & 1.84 (1.75 & 0.25H, each d, each J=1.0 Hz), 1.30 & 1.29 (9H,
each d, J=6.5 & 7.0 Hz).
Example 8
(a) N-(2-hydroxyethyl)-N-2-(2-propyldithio)-1-propenyl]formamide
In a solution of sodium hydroxide (1.3 g) in water (50 ml) was dissolved
Intermediate 5 (3.4 g). To this solution was added sodium
2-propylthiosulfate (ca. 7 g) in the form of powder, and the mixture was
stirred at room temperature for 0.5 hour. The reaction was extracted with
chloroform. After the chloroform layer was dried with anhydrous sodium
sulfate, the solvent was evaporated under reduced pressure. The residue
obtained was purified by silica gel column chromatography (CHCl.sub.3
:MeOH=50:1.fwdarw.35:1).
Yield: 1.63 g.
.sup.1 H-NMR (in CDCl.sub.3, 6:1 isomeric mixture): .delta.8.18 & 8.10
(0.85 & 0.15H, each s), 6.05 & 5.99 (0.85 & 0.15H, each s), 3.79 & 3.70
(1.7 & 0.3H, each m), 3.62 (2H, t), 2.97 (1H, m), 2.35 (1H, broad), 2.19 &
2.17 (2.6 & 0.4H, each s), 1.30 & 1.29 (6H, each d).
(b)
N-[2-[2-[(5-dimethylaminonaphthylsulfonyl)amino]propionyloxy]ethyl-N-[2-(2
-propyldithio)-1-propenyl]formamide
Compound C (2.2 g), the compound in the above step (a) (1.60 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (1.70 g) was added to the solution. The mixture
was then stirred at room temperature for about 10 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.10:1.5).
Yield: 2.19 g.
.sup.1 H-NMR (in CDCl.sub.3, 6:1 isomeric mixture): .delta.8.54 & 8.29
(each 1H, each d), 8.24 (1H, dd), 8.06 & 7.87 (0.86 & 0.14H, each s),
7.62-7.50 (2H, m), 7.20 (1H, d), 5.97 & 5.90 (0.14 & 0.86H, each d, each
J=1.5 Hz), 5.45 & 5.33 (0.86 & 0.14H, each d, each J=8.5 Hz), 4.00-3.94
(2H, m), 3.87 (1H, m), 3.49 (2H, m), 2.94 (1H, m), 2.89 (6H, s), 2.14 &
2.13 (3H, each d, each J=1.0 Hz), 1.30 & 1.29 (9H, each d, J=7.5 & 6.5
Hz).
Example 9
(a)
N,N'-{dithiobis[2-methyl-2,1-ethenediyl]}bis{N-[(2-hydroxy)ethyl]formamide
In a solution of sodium hydroxide (1.68 g) in water (20 ml) was dissolved
Intermediate 5 (4.5 g). To this solution was added portionwise an aqueous
iodine-potassium iodide solution separately prepared (0.5M). While for
some time from initiating the addition, the brown color of iodine
immediately disappeared as soon as the aqueous iodine-potassium iodide
solution was added. The color faded gradually and finally the solution
remained pale brown. At this time, the addition of the aqueous
iodine-potassium iodide solution was stopped (the reaction has a pH of
about 7 at this time). The reaction was saturated with sodium sulfate and
extracted with chloroform. After the chloroform layer was dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :ethanol=40:1.fwdarw.15:1).
Yield: 2.17 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1:1 isomeric mixture): 8 8.16, 8.15 & 8.09
(0.17, 0.66 & 0.17H, each s), 6.16, 6.14 & 6.07 (0.66 & 0.17 & 0.17H, each
s), 3.78 & 3.71 (1.8 & 0.17H, each m), 3.62 (2H, m), 2.58 & 2.40 (0.17 &
0.83H, each t), 2.14 (3H, m).
.sup.1 H-NMR (in DMSO-d.sub.6, 8:5:5:2 isomeric mixture): .delta.8.052,
8.046 & 8.039 & 8.032 (1H, each s), 6.43, 6.37, 6.19 & 6.16 (0.4, 0.25,
0.25 & 0.1H, each d, each J=1.5 Hz), 4.82 & 4.77 (0.33 & 0.67H, t & m),
3.53-3.42 (4H, m), 2.03 (3H, m).
(b)
N,N'-{dithiobis[2-methyl-2,1-ethenediyl]}bis{N-[2-[3-[(5-dimethylaminonaph
thylsulfonyl)amino]propionyloxy]ethyl]formamide
Compound B (4.4 g), the compound in the above step (a) (2.12 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (40 ml).
Dicyclohexylcarbodiimide (1.70 g) was added to the solution. And then the
mixture was stirred at room temperature for about 16 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.10:3) to give the title
compound.
Yield: 1.66 g.
.sup.1 H-NMR (in CDCl.sub.3): .delta.8.54, 8.30 & 8.23 (each 1H, each d),
8.1 (1H, m), 7.58-7.50 (2H, m), 7.18 (1H, d), 6.33-6.08 (1H, m), 5.82 (1H,
broad m), 4.12 (2H, m), 3.67 (2H, m), 3.14 (2H, m), 2.89 (6H, s), 2.50
(2H, m), 2.08 (3H, m).
Example 10
(a) N-(2-hydroxyethyl)-N-[1-methyl-2-(2-Propyldithio)vinyl]-acetamide
In a solution of sodium hydroxide (1.3 g) in water (50 ml) was dissolved
Intermediate 6 (3.57 g). To this solution was added sodium
2-propylthiosulfate (ca. 6 g) in the form of powder, and the mixture was
stirred at room temperature for about 0.5 hour. The reaction mixture was
extracted with chloroform. After the chloroform layer was dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=40:1.fwdarw.35:1).
Yield: 2.33 g.
.sup.1 H-NMR (in CDCl.sub.3): .delta.6.08 (1H, d, J=1.0 Hz), 3.81 (2H,
broad), 3.73 & 3.46 (each 1H, each broad), 3.30 (1H, t), 3.05 (1H, m),
2.10 (3H, s), 1.94 (3H, J=1.0 Hz), 1.32 (6H, d, J=6.5 Hz).
(b)
N-[2-[2-[(5-dimethylaminonaphthylsulfonyl)amino]propionyloxy]ethyl]-N-[1-m
ethyl-2-(2-propyldithio)vinyl]acetamide
Compound C (3.0 g), the compound in the above step (a) (2.3 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (1.96 g) was added to the solution. And then the
mixture was stirred at room temperature for about 18 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.9:1) to give the title
compound.
Yield: 3.85 g.
.sup.1 H-NMR (in CDCl.sub.3): .delta.8.54 & 8.30 (each 1H, each d), 8.23
(1H, dd), 7.59 & 7.51 (each 1H, each dd), 7.19 (1H, d), 6.05 (1H, broad
s), 5.50 (1H, broad d), 3.99 (2H, m), 3.90 (1H, broad), 3.60 & 3.31 (each
1H, each broad), 3.03 (1H,.m), 2.88 (6H, s), 2.03 (3H, s), 1.84 (3H, s),
1.30 & 1.29 (9H, each d).
Example 11
(a)
N-(2-hydroxyethyl)-N-[4-hydroxy-1-methyl-2-(2-propyldithio)-1-butenyl]form
amide
In a solution of sodium hydroxide (1.3 g) in water (50 ml) was dissolved
Intermediate 7 (4.02 g). After this solution was saturated with sodium
sulfate, sodium 2-propylthiosulfate (ca. 7 g) was added in the form of
powder. The mixture was then stirred at room temperature for about 15
minutes. The reaction mixture was extracted with chloroform. After the
chloroform layer was dried with anhydrous sodium sulfate, the solvent was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=40:1.fwdarw.20:1)
to give the title compound.
Yield: 3.90 g.
.sup.1 H-NMR (in CDCl.sub.3, 2:1 isomeric mixture): .delta.8.03 & 8.02 (1H,
each s), 3.85-3.78 (4H, m), 3.72 & 3.65 (2H, each m), 3.00-2.98 (3H, m),
2.05 & 1.99 (2 & 1H, each s), 1.28 (6H, d, J=7.0 Hz).
.sup.1 H-NMR (in DMSO-d.sub.6, 7:3 isomeric mixture): .delta.7.98 & 7.87
(0.3 & 0.7H, each s), 4.82, 4.74, 4.69 & 4.65 (0.3, 0.7, 0.7 & 0.3H, each
t), 3.52-3.45 (4H, m), 3.42 (2H, t), 2.96 (1H, m), 2.73 (2H, t), 1.99 &
1.92 (0.9 & 2.1H, each s), 1.21 & 1.20 (6H, each d, J=6.5 & 5.0 Hz).
(b)
N-[2-[3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyl)oxy]ethyl-N-[4-[
3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyl]oxy-1-methyl-2-(2-propy
ldithio)-1-butenyl]formamide
Compound B (4.28 g), the compound in the above step (a) (1.80 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
Dicyclohexylcarbodiimide (3.00 g) was added to the solution. And then the
mixture was stirred at room temperature for about 12 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.10:2) to give the title
compound.
Yield: 3.96 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.54 (2H, dd),
8.30-8.22 (4H, m), 7.95 & 7.94 (0.8 & 0.2H, each s), 7.18 (2H, dd), 5.82,
5.68 5.47 & 5.40 (0.2, 0.8, 0.8 & 0.2H, each broad t), 4.15 & 4.07 (3.6 &
0.4H, each m), 3.66 & 3.59 (1.6 & 0.4H, each broad), 3.16 (4H, m),
2.94-2.88 (15H, m), 2.50 (4H, m), 1.98 & 1.94 (2.4 & 0.6H, each s), 1.25
(6H, d).
Example 12
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-propyl
)dithio]-1-butenyl]formamide
In a solution of sodium hydroxide (2.0 g) in water (50 ml) was dissolved
Intermediate 8 (9.2 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 2-propylthiosulfate (8 g) in the form of
powder was added to the aqueous solution. Immediately after the addition,
a pale brown oil product was deposited. The reaction mixture was extracted
with ethyl acetate (100 ml). The ethyl acetate layers were combined, dried
over anhydrous sodium sulfate and evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography to give
the title compound of which amino group was protected with Boc group.
Yield: 9.07 g.
.sup.1 H-NMR (in CD.sub.3 OD, 5:2 isomeric mixture): .delta.8.00 & 7.93
(0.29 & 0.71H, each s), 3.72 (2H, t), 3.53 (2H, broad m), 3.23 (2H, t),
2.89 (2H, t), 2.67 (2H, t), 2.06 & 2.01 (2.14 & 0.86H, each s), 1.67 (2H,
m), 1.43 (9H, s), 0.99 (3H, m).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-[(1-propyl)dithio)-1-butenyl]formamide
hydrochloride
In a solution of the compound in the above step (a) (9.07 g) in
tetrahydrofuran (50 ml) was dissolved carbodiimidazole (4.05 g) in a water
bath at 40.degree. C. To this solution was added Compound A (7.6 g), and
the mixture was stirred at room temperature for 24 hours. After insolubles
were removed by filtration, the filtrate was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography to give the title compound of which amino group was
protected with Boc group.
Yield: 15.3 g.
.sup.1 H-NMR (in CD.sub.3 OD, 3:1 isomeric mixture): .delta.8.56, 8.33,
8.19 (each 1H, each d), 7.97 & 7.87 (0.25 & 0.75H, each s), 7.58 (2H, m),
7.29 (1H, d), 4.14 (2H, t), 3.49 & 3.19 (each 2H, each broad t), 3.10 (2H,
t), 2.95-2.88 (4H, m), 2.89 (6H, s), 2.65 (2H, m), 2.00 & 1.95 (2.1 &
0.9H, each s), 1.64 (2H, m), 1.41 (9H, s), 0.97 (3H, m).
The Boc-protected derivative (15.3 g) was added with trifluoroacetic acid
(40 ml) and dissolved under ice-cooling. And then the mixture was left
standing at room temperature for about 10 minutes. The trifluoroacetic
acid solution was slowly added dropwise to an aqueous sodium hydrogen
carbonate suspension (200 ml). Yellow fluorescent viscous powder was
deposited. The mixture was extracted with ethyl acetate (200 ml). The
ethyl acetate layer was washed with water and dried over anhydrous sodium
sulfate. The solvent was then evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=45:1.fwdarw.20:1) to give 11.12 g of a residue. It
was dissolved in ethanol and concentrated after 1N hydrochloric acid (18.6
ml) was added. The residue having ethanol added thereto was concentrated
again and added dropwise to a mixed solvent of ether:hexane=2:1 (400 ml).
Pale yellow powdery solid obtained was collected by filtration and dried
under reduced pressure to give the title compound.
Yield: 9.17 g.
.sup.1 H-NMR (in CD.sub.3 OD, 10:1 isomeric mixture): .delta.8.56 & 8.51
(each 1H, each d), 8.24 (1H, dd), 7.96 & 7.89 (0.9 & 0.1H, each s),
7.70-7.67 (2H, m), 7.56 (1H, broad d), 4.20 (2H, t), 3.71 (2H, t), 3.16
(2H, t), 3.12 (2H, t), 3.09 (6H, s), 2.98 (2H, t), 2.91 (2H, t), 2.70 (2H,
t), 2.08 & 2.00 (2.7 & 0.3H, each s), 1.67 (2H, q), 0.99 (3H, t).
Example 13
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(2-propyl
)dithio]-1-butenyl]formamide
In a solution of sodium hydroxide (2.0 g) in water (30 ml) was dissolved
Intermediate 8 (9.4 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 2-propylthiosulfate (9.0 g) in the form
of powder was added to the aqueous solution. Immediately after the
addition, a pale yellow oil product was deposited. The reaction mixture
was extracted with chloroform (100 ml). The aqueous layer was further
extracted with chloroform (100 ml). The chloroform layers were combined,
dried with anhydrous sodium sulfate and evaporated under reduced pressure.
The residue was purified by silica gel column chromatography to give the
title compound of which amino group was protected with Boc group.
Yield: 6.81 g.
.sup.1 H-NMR (in CD.sub.3 OD, 5:2 isomeric mixture): .delta.8.00 & 7.96
(0.28 & 0.72H, each s), 3.70 (2H, t), 3.54 & 3.25 (each 2H, each broad t),
2.96 (1H, m), 2.89 (2H, t), 2.06 & 2.01 (2.1 & 0.9H, each s), 1.43 (9H,
s), 1.28 (6H, d).
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.05 & 7.97
(0.75 & 0.25H, each s), 3.80 (2H, t), 3.78 (2H, t), 3.57 (2H, broad), 3.31
(2H, broad t), 2.95 (1H, m), 2.88 (2H, t), 2.03 & 1.99 (2.25 & 0.75H, s),
1.45 & 1.43 (9H, each s), 1.28 (6H, d).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-[(2-propyl)-dithio)-1-butenyl]formamide
hydrochloride
In a solution of the compound in the above step (a) (3.8 g) in
tetrahydrofuran (20 ml) was dissolved carbonyldiimidazole (1.7 g) in the
form of powder in a water bath at 40.degree. C. To this solution was added
Compound A (2.95 g), and the mixture was stirred at room temperature for
about 5 hours. After insolubles were removed by filtration, the filtrate
was evaporated under reduced pressure. The residue obtained was purified
by silica gel column chromatography to give the Boc-protected derivative
of the title compound (CHCl.sub.3 :methanol=80:1.fwdarw.50:1).
Yield: 6.68 g.
.sup.1 H-NMR (in CD.sub.3 OD, 7:3 isomeric mixture): .delta.8.56, 8.32,
8.18 (each 1H, each d), 7.99 & 7.91 (0.3 & 0.7H, each s), 7.59 (2H, m),
7.27 (1H, d), 4.12 (2H, t), 3.51 (2H, broad), 3.21 (2H, broad), 3.10 (2H,
t), 2.96-2.89 (5H, m), 2.88 (6H, s), 2.00 & 1.94 (2.1 & 0.9H, each s),
1.41 (9H, s), 1.26 (6H, d).
The Boc-protected derivative (3.3 g) was added with trifluoroacetic acid
(15 ml) and dissolved under ice-cooling. The mixture was then left
standing at room temperature for about 10 minutes. The trifluoroacetic
acid solution was slowly added dropwise to an aqueous sodium hydrogen
carbonate suspension (100 ml). Yellow fluorescent viscous powder was
deposited. The mixture was extracted with ethyl acetate (100 ml). After
the ethyl acetate layer was washed with water, it was partitioned with 1N
hydrochloric acid (50 ml). The aqueous layer was adjusted to a pH of 9
with sodium sulfate and partitioned with chloroform (100 ml). After the
chloroform layer was dried with anhydrous sodium sulfate, the solvent was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=40:1.fwdarw.20:1)
to give 2.39 g of a residue. It was dissolved in ethanol (150 ml) and
concentrated after 1N hydrochloric acid (4.0 ml) was added. The residue
having ethanol added thereto was concentrated again and added dropwise to
ether (200 ml). Pale yellow powdery solid obtained was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 2.24 g.
.sup.1 H-NMR (in CD.sub.3 OD, 10:1 isomeric mixture): .delta.8.57 (1H, d),
8.36 (1H, d), 8.20 (1H, d), 8.08 & 7.99 (0.1 & 0.9H, each s), 7.63-7.59
(2H, m), 4.18 (2H, t), 3.72 (2H, t), 3.18 (2H, t), 3.12 (2H, t), 3.02-2.96
(3H, m), 2.94 (6H, s), 2.90 (2H, t), 2.70 & 2.00 (3H, each s), 1.28 (6H,
d).
Example 14
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(2-butyl)
dithio]-1-butenyl]formamide
In a solution of sodium hydroxide (2.0 g) in water (50 ml) was dissolved
Intermediate 8 (9.2 g). Sodium 2-butylthiosulfate (8 g) in the form of
powder was added to the aqueous solution. Immediately after the addition,
a pale brown oil product was deposited. After the reaction mixture was
partitioned with ethyl acetate (200 ml), the ethyl acetate layer was dried
over anhydrous sodium sulfate and evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography (only
CHCl.sub.3 .fwdarw.CHCl.sub.3 :methanol=40:1) to give the title compound
of which amino group was protected with Boc group.
Yield: 4.1 g.
.sup.1 H-NMR (in CD.sub.3 OD, 3:1 isomeric mixture): .delta.8.00 & 7.96
(0.25 & 0.75H, each s), 3.70 (2H, t), 3.53 (2H, broad t), 3.24 (2H, broad
t), 2.90 (2H, t), 2.76 (1H, m), 2.06 & 2.01 (2.1 & 0.9H, each s), 1.67 &
1.52 (each 1H, each m), 1.43 (9H, s), 1.27 (3H, d), 0.98 (3H, d).
(b)
N-(2-aminoethyl)-N[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamino
carbonyloxy]-2-[(2-butyl)dithio)-1-butenyl]formamide hydrochloride
In a solution of the compound in the above step (a) (4.1 g) in
tetrahydrofuran (30 ml) was dissolved carbonyldiimidazole (1.8 g) in the
form of powder, and the mixture was left standing at room temperature for
1 hour. To this solution was added Compound A (2.95 g), and the mixture
was stirred at room temperature for about hours. After insolubles were
removed by filtration, the filtrate was evaporated under reduced pressure.
The residue obtained was purified by silica gel column chromatography to
give the Boc-protected derivative of the title compound (CHCl.sub.3
:methanol=80:1.fwdarw.50:1).
Yield: 2.92 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 4:1 isomeric mixture): .delta.8.46 (1H, d),
8.26 (1H, d), 8.09 (1H, d), 7.95 (1H, broad s), 7.79 (1H, s), 7.63-7.57
(2H, m), 7.26 (1H, d), 7.01 (1H, broad), 6.79 (1H, broad), 4.01 & 3.95
(1.5 & 0.5H, each t), 3.36 (2H, m), 3.04 (2H, q), 2.98 (2H, q), 2.83 (6H,
s), 2.8-2.72 (5H, m), 1.92 & 1.87 (2.4 & 0.6H, s), 1.57 & 1.44 (each 1H,
each m), 1.36 (9H, s), 1.18 (3H, d), 0.89 (3H, t).
The Boc-protected derivative (2.8 g) was ice-cooled, added with
trifluoroacetic acid (30 ml) and dissolved under ice-cooling. And then the
mixture was left standing at room temperature for about 15 minutes. The
trifluoroacetic acid solution was slowly added dropwise to an aqueous
sodium hydrogen carbonate suspension (150 ml). Yellow fluorescent viscous
powder was deposited. The mixed solution was extracted with chlroform (100
ml). After the chloroform layer was washed with water and dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=50:1.fwdarw.20:1) to give 2.15 g of a
foam. After the foam was dissolved in ethanol (150 ml) and added with 1N
hydrochloric acid (3.5 ml), the mixture was concentrated. The residue
having ethanol added thereto was concentrated again and dissolved in
chloroform (20 ml). The solution was added dropwise to ether (250 ml).
Pale yellow powdery solid obtained was collected by filtration and dried
under reduced pressure to give the title compound.
Yield: 1.82 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 6:1 isomeric mixture): .delta.8.49, 8.30 &
8.10 (each 1H, each d), 8.05-7.97 (4H, broad), 7.85 (1H, s), 7.61 (2H, m),
7.29 (1H, d), 7.11 (1H, broad), 4.02 (2H, t), 3.61 (2H, t), 2.99 (2H, q),
2.88 (2H, broad), 2.85 (6H, s), 2.85-2.74 (5H, m), 1.96 & 1.89 (2.6 &
0.4H, each s), 1.55 & 1.46 (each 1H, each m), 1.19 (3H, d), 0.90 (3H, t).
Example 15
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-pentyl
)dithio]-1-butenyl]formamide
In a solution of sodium hydroxide (4.0 g) in water (50 ml) was dissolved
Intermediate 8 (18.4 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 1-pentylthiosulfate (20 g) in the form
of powder was added to the aqueous solution. Immediately after the
addition, a pale brown oil product was deposited. The reaction mixture was
partitioned with chloroform (100 ml). Then the aqueous layer was further
partitioned with chloroform (100 ml). These chloroform layers were
combined, dried with anhydrous sodium sulfate and evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography to give the title compound of which amino group was
protected with Boc group.
Yield: 18.22 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 3:1 isomeric mixture): .delta.7.94 & 7.83
(0.25 & 0.75H, each s), 6.92 & 6.83 (0.25 & 0.75H, each broad), 4.71 &
4.68 (0.75 & 0.25H, each t), 3.52 & 3.47 (1.5 & 0.5H, q & m), 3.36 (2H,
broad), 3.04 (2H, t), 2.74 (2H, t), 2.65 (2H, t), 1.98 & 1.91 (2.25 &
0.75H, each s), 1.57 (2H, m), 1.37 (9H, s), 1.28 (4H, m), 0.85 (3H, m).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-[(1-pentyl)dithio)-1-butenyl]formamide
hydrochloride
In a solution of the compound in the above step (a) (6.1 g) in
tetrahydrofuran (25 ml) was dissolved carbonyldiimidazole (2.56 g) in the
form of powder in a water bath at 40.degree. C. To this solution was added
Compound A (4.7 g), and the mixture was stirred at room temperature for
about 10 hours. After insolubles were removed by filtration, the filtrate
was evaporated under reduced pressure. The residue obtained was purified
by silica gel column chromatography to give the Boc-protected derivative
of the title compound (CHCl.sub.3 :methanol=80:1.fwdarw.40:1).
Yield: 5.75 g.
.sup.1 H-NMR (in CD.sub.30 D, 3:1 isomeric mixture): .delta.8.56, 8.31,
8.18 (each 1H, each d), 7.89 & 7.87 (0.25 & 0.75H, each s), 7.58 (2H,q),
7.28 (1H, d), 4.14 (2H, t), 3.49 (2H, broad), 3.19 (2H, t), 3.10 (2H, t),
2.95-2.88 (4H, m), 2.88 (6H, s), 2.67 (2H, t), 2.00 & 1.95 (2.25 & 0.75H,
t), 1.62 (2H, m), 1.41 (9H, s), 1.33 (4H, m), 0.90 (3H, t).
The Boc-protected derivative (5.66 g) was ice-cooled, added with
trifluoroacetic acid (30 ml) and dissolved under ice-cooling. And then the
mixture was left standing at room temperature for about 40 minutes. The
trifluoroacetic acid solution was slowly added dropwise to an aqueous
sodium hydrogen carbonate suspension (150 ml). Yellow fluorescent viscous
powder was then deposited. The mixed solution was partitioned with
chlroform (100 ml). After the chloroform layer was washed with water and
dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography to give 4.54 g of a residue. After the residue was
dissolved in ethanol (150 ml) and added with 1N hydrochloric acid (7.3
ml), the mixture was concentrated. The residue having ethanol added
thereto was concentrated again and dissolved in trichloromethane (15 ml).
The solution was added dropwise to ether (150 ml). Pale yellow viscous
solid obtained was solidified in hexane. The powdery solid obtained was
collected by filtration and dried under reduced pressure to give the title
compound.
Yield: 4.55 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 10:1 isomeric mixture): .delta.8.48, 8.28 &
8.09 (each 1H, each d), 8.04-7.96 (3H, broad), 7.82 (1H, s), 7.61 (2H, m),
7.28 (1H, d), 7.14 (1H, t), 4.03 (2H, t), 3.59 (2H, broad), 2.99 (2H, q),
2.90-2.78 (12H, m), 2.67 (2H, t), 1.97 & 1.90 (2.7 & 0.3H, s), 1.55 (1H,
m), 1.32 -1.24 (4H, m), 0.85 (3H, m).
Example 16
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-methyl
butyl)dithio]-1-butenyl]formamide
In a solution of sodium hydroxide (4.0 g) in water (120 ml) was dissolved
Intermediate 8 (18.4 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 1-methylbutylthiosulfate (20 g) in the
form of powder was added to the aqueous solution. Immediately after the
addition, a yellowish brown viscous oil product was floated. After the
reaction mixture was partitioned with chloroform, the oily components were
extracted and washed with water. The combined chloroform layers were dried
with anhydrous sodium sulfate and evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :MeOH=40:1.fwdarw.30:1) to give the title compound of which
amino group was protected with Boc group.
Yield: 18.92 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 4:1 isomeric mixture): .delta.7.95 & 7.86
(0.2 & 0.8H, each s), 6.92 & 6.83 (0.2 & 0.8H, each broad), 4.71 & 4.66
(0.8 & 0.2H, each t), 3.51 & 3.46 (1.6 & 0.4H, each q), 3.37 (2H, broad),
3.06 (2H, q), 2.81 (1H, m), 2.74 (2H, t), 1.98 & 1.90 (2.4 & 0.6H, each
s), 1.54 (1H, m), 1.37 (10H, m & s), 1.20 (3H, d), 0.87 (3H, t).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-[(1-methylbutyl)dithio)-1-butenyl]formamide
hydrochloride
In a solution of the compound in the above step (a) (4.3 g) in
tetrahydrofuran (15 ml) was dissolved carbonyldiimidazole (1.8 g) in the
form of powder, and the mixture was heated in a water bath at 50.degree.
C. for about 20 minutes. To this solution was added Compound A (3.3 g),
and the mixture was stirred at room temperature for about 12 hours. After
insolubles and deposits were removed by filtration, the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography to give the Boc-protected derivative of
the title compound (CHCl.sub.3 :methanol=80:1.fwdarw.60:1).
Yield: 3.40 g.
.sup.1 H-NMR (in CD.sub.3 OD, 5:2 isomeric mixture): .delta.8.56, 8.31,
8.18 (each 1H, each d), 7.98 & 7.90 (0.29 & 0.71H, each d), 7.58 (2H, q),
7.27 (1H, d), 4.12 (2H, t), 3.50 (2H, m), 3.20 (2H, t), 3.10 (2H, t), 2.95
& 2.91 (4H, m), 2.88 (6H, s), 2.80 (1H, m), 2.00 & 1.94 (1.14 & 0.86H,
each s), 1.58 (1H, m), 1.48-1.38 (12H, m), 1.25 (3H, d), 0.91 (3H, t).
The Boc-protected derivative (3.22 g) was ice-cooled, added with
trifluoroacetic acid (20 ml) and dissolved under ice-cooling, and the
mixture was left standing at room temperature for about 40 minutes. The
trifluoroacetic acid solution was slowly added dropwise to an aqueous
sodium hydrogen carbonate suspension (150 ml). A yellow fluorescent
viscous product was then deposited. This mixed solution was partitioned
with chloroform (100 ml). After the chloroform layer was washed with water
and dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography to give 2.70 g of a residue. After the residue was
dissolved in ethanol (150 ml) and added with 1N hydrochloric acid (7.3
ml), the mixture was concentrated. The residue having ethanol added
thereto was concentrated again and dissolved in chloroform (15 ml). The
solution was added dropwise to ether (150 ml). Pale yellow powdery solid
obtained was collected by filtration and dried under reduced pressure to
give the title compound.
Yield: 2.48 g.
.sup.1 H-NMR (in CD.sub.3 OD, 9:1 isomeric mixture): .delta.8.57, 8.31 &
8.18 (each 1H, each d), 8.07 & 7.99 (0.1 & 0.9H, each s), 7.58 (2H, q),
7.28 (1H, d), 4.18 (2H, t), 3.71 (2H, t), 3.18 (2H, t), 3.12 (2H, t), 2.98
(2H, t), 2.92-2.84 (9H, m), 2.08 & 2.00 (2.7 & 0.3H, each s), 1.58 (1H,
m), 1.50-1.40 (3H, m), 1.28 (3H, d), 0.92 (3H, t).
Example 17
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-2-[(3-pentyl)dithio]-
1-butenyl]formamide
In a solution of sodium hydroxide (4.0 g) in water (100 ml) was dissolved
Intermediate 8 (18.4 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 3-pentylthiosulfate (20 g) in the form
of powder was added to the aqueous solution. Immediately after the
addition, a yellowish brown viscous oil product was deposited. After the
reaction mixture was partitioned with chloroform, the oily components were
extracted and washed with water. The combined chloroform layers were dried
with anhydrous sodium sulfate and evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=80:1.fwdarw.30:1) to give the title compound of
which amino group was protected with Boc group.
Yield: 15.3 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 4:1 isomeric mixture): .delta.7.95 & 7.85
(0.2 & 0.8H, each s), 6.86 & 6.82 (0.2 & 0.8H, each broad), 4.71 & 4.67
(0.8 & 0.2H, each t), 3.52 & 3.47 (1.6 & 0.4H, each q), 3.37 (2H, broad),
3.05 (2H, q), 2.75 (2H, t), 2.58 (1H, m), 1.98 & 1.90 (2.4 & 0.6H, each
s), 1.54 (4H, m), 1.37 (9H, s), 0.91 (6H, m).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-[(3-pentyl)dithio)-1-butenyl]formamide
hydrochloride
In a solution of the compound in the above step (a) (6.1 g) in
tetrahydrofuran (25 ml) was dissolved carbonyldiimidazole (2.6 g) in the
form of powder in a water bath at 40.degree. C. To this solution was added
Compound A (4.7 g), and the mixture was stirred at room temperature for
about 13 hours. After insolubles and deposits were removed by filtration,
the filtrate was evaporated under reduced pressure. The residue obtained
was purified by silica gel column chromatography to give the Boc-protected
derivative of the title compound (CHCl.sub.3 :methanol=80:1.fwdarw.40:1).
Yield: 6.54 g.
.sup.1 H-NMR (in CD.sub.3 OD, 4:1 isomeric mixture): .delta.8.56, 8.32,
8.18 (each 1H, each d), 7.90 & 7.89 (0.2 & 0.8H, each s), 7.58 (2H, q),
7.28 (1H, d), 4.13 (2H, t), 3.49 (2H, broad), 3.20 (2H, t), 3.10 (2H, t),
2.95 & 2.91 (4H, m), 2.88 (6H, s), 2.57 (1H, m), 2.00 & 1.94 (2.4 & 0.6H,
each s), 1.61 (4H, m), 1.43 & 1.41 (9H, s), 0.96 (6H, t).
The Boc-protected derivative (6.0 g) was ice-cooled, added with
trifluoroacetic acid (30 ml) and dissolved under ice-cooling. And then the
mixture was left standing at room temperature for about 15 minutes. The
trifluoroacetic acid solution was slowly added dropwise to an aqueous
sodium hydrogen carbonate suspension (200 ml). A yellow fluorescent
viscous product was then deposited. This mixed solution was partitioned
with chloroform. After the chloroform layer was washed with water and
dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 +CHCl.sub.3 :MeOH=30:1.fwdarw.15:1) to give
7.20 g of a residue. After the residue was dissolved in 2-propanol (200
ml) and added with 1N hydrochloric acid (11.5 ml), the mixture was
concentrated. After the residue was dissolved in chloroform, the solution
was added dropwise to ether (150 ml). As the deposit was viscous, the
ether layer was discarded, and then the residue was solidified by the
addition of hexane. Pale yellow powdery solids obtained were collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 7.02 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.8.47, 8.27 &
8.09 (each 1H, each d), 8.04-7.97 (4H, broad), 7.86 & 7.83 (0.1 & 0.9H),
7.61 (2H, m), 7.27 (1H, d),7.12 (1H, broad), 4.04 (2H, t), 3.59 (2H, broad
t), 2.99 (2H, broad q), 2.92-2.78 (12H, m), 2.60 (1H, m), 1.96 (3H, s),
1.53 (4H, m), 0.90 (6H, t).
Example 18
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-2-cyclohexyldithio-1-
methyl-1-butenyl]formamide
To a solution of sodium hydroxide (2.0 g) in water (80 ml) was added
Intermediate 8 (9.2 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium cyclohexylthiosulfate (10 g) in the form
of powder was added to the aqueous solution. Immediately after the
addition, a pale yellow oil product was deposited. After the reaction
mixture was partitioned with chloroform (100 ml), the aqueous layer was
further partitioned with chloroform (100 ml). The combined chloroform
layers were dried with anhydrous sodium sulfate and evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography to give the title compound of which amino group was
protected with the Boc group.
Yield: 1.81 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 3:1 isomeric mixture): .delta.7.95 & 7.85
(0.25 & 0.75H, each s), 6.92 & 6.82 (0.25 & 0.75H, each broad), 4.68 &
4.65 (0.75 & 0.25H, each broad t), 3.51 & 3.46 (1.5 & 0.5H, each q), 3.38
(2H, broad), 3.07 (2H, broad q), 2.76-2.68 (3H, m), 1.98 (3H, s), 1.90,
1.68 & 1.53 (5H, m), 1.37 (9H, s), 1.28-1.14 (5H, m).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-cyclohexyldithio-1-butenyl]formamide hydrochloride
In a solution of the compound prepared in the above step (a) (1.77 g) in
tetrahydrofuran (15 ml) was dissolved carbonyldiimidazole (0.72 g) in the
form of powder in a water bath at 40.degree. C. To this solution was added
Compound A (1.5 g), and the mixture was stirred at room temperature for
about 15 hours. After insolubles and deposits were removed by filtration,
the filtrate was evaporated under reduced pressure. The residue obtained
was purified by silica gel column chromatography to give the Boc-protected
derivative of the title compound (CHCl.sub.3 :MEOH=80:1.fwdarw.30:1).
Yield: 2.71 g.
.sup.1 H-NMR (in CD.sub.3 OD): .delta.8.56, 8.32 & 8.19 (each 1H, each d),
7.88 (1H, d), 7.58 (2H, q), 7.27 (1H, d), 4.12 (2H, t), 3.30 (2H, broad),
3.22 (2H, t), 3.10 (2H, t), 2.96 & 2.90 (4H, m), 2.88 (6H, s), 2.70 (1H,
m), 1.99 (3H, s), 1.98-1.94 (2H, m), 1.75 (2H, m), 1.60 (1H, m), 1.41 (9H,
s), 1.34-1.28 (5H, m).
The Boc-protected derivative (2.64 g) was ice-cooled, added with
trifluoroacetic acid (30 ml) and dissolved under ice-cooling. And then the
mixture was left standing in a water bath at 50.degree. C. for about 15
minutes. The trifluoroacetic acid solution was slowly added dropwise to an
aqueous sodium hydrogen carbonate suspension (150 ml). A yellow
fluorescent viscous product was then deposited. This mixed solution was
partitioned with chlroform (100 ml). After the chloroform layer was washed
with water and dried with anhydrous sodium sulfate, the solvent was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.15:1)
to give 1.76 g of a residue. After the residue was dissolved in ethanol
(150 ml) and added with 1N hydrochloric acid (2.8 ml), the mixture was
concentrated. The residue was added with ethanol and concentrated again.
The residue was dissolved in chloroform (10 ml) and added dropwise to
ether (150 ml). The pale yellow powdery product was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 1.24 g.
.sup.1 H-NMR (in CD.sub.3 OD, 12:1 isomeric mixture): .delta.8.57, 8.39 &
8.21 (each 1H, each d), 8.07 & 7.99 (0.08 & 0.92H, each s), 7.62 (2H, q),
7.38 (1H, d), 4.17 (2H, t), 3.72 (2H, t), 3.19 (2H, t), 3.12 (2H, t), 2.98
(2H, t), 2.96 (6H, s), 2.92 (2H, t), 2.76 (1H, m), 2.07 (3H, s), 1.98 (2H,
m), 1.77 (2H, m), 1.62 (1H, m), 1.38-1.24 (5H, m).
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.8.48, 8.28 & 8.09 (each 1H, each d),
7.99 (4H, broad), 7.85 (1H, s), 7.61 (2H, m), 7.28 (1H, d), 7.10 (1H,
broad), 4.02 (2H, t), 3.60 (2H, t), 2.99 (2H, q), 2.89 (2H, t), 2.85 (6H,
s), 2.84-2.79 (4H,m), 2.74 (1H, m), 1.96 (3H, s), 1.89 (2H, m), 1.67 (2H,
m), 1.53 (1H, m), 1.30-1.15 (5H, m).
Example 19
(a)
N-[2-(tert-butyloxycarbonylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-dodecy
l)dithio]-1-butenyl]formamide
To a solution of sodium hydroxide (2.0 g) in water (70 ml) was added
Intermediate 8 (9.2 g), and the mixture was left standing at room
temperature for 5 minutes. Sodium 1-dodecyl-thiosulfate (6 g) in the form
of powder was added to the aqueous solution. Sonication was conducted
because of the low dissolution rate of the powder. Along with the
dissolution of it, a yellowish brown amorphous product was deposited.
After the reaction was partitioned with chloroform (100 ml), the aqueous
layer was further partitioned with chloroform (100 ml). The combined
chloroform layers were dried with anhydrous sodium sulfate and evaporated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography to give the title compound of which amino group was
protected with Boc group.
Yield: 4.11 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 4:1 isomeric mixture): 7.95 & 7.83 (0.2 &
0.8H, each s), 6.88 & 6.79 (0.2 & 0.8H, each broad), 4.68 (1H, broad t),
3.53 & 3.48 (1.6 & 0.4H, each q), 3.36 (2H, broad), 3.05 (2H, broad), 2.74
(2H, t), 2.65 (2H, t), 1.98 & 1.91 (2.4 & 0.63H, each s), 1.98 & 1.91 (2.4
& 0.63H, each s), 1.56 (2H, m), 1.37 (9H, s), 1.32-1.24 (18H, m), 0.86
(3H, t).
(b)
N-(2-aminoethyl)-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylamin
ocarbonyloxy]-1-methyl-2-dodecyldithio-1-butenyl]formamide hydrochloride
In a solution of the compound prepared in the above step (a) (4.07 g) in
tetrahydrofuran (30 ml) was dissolved carbonyldiimidazole (1.4 g) in the
form of powder, and the mixture was left standing at room temperature for
about 4 hours. To this solution was added Compound A (2.6 g), and the
mixture was stirred at room temperature for about 16 hours. After
insolubles and deposits were removed by filtration, the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography to give the Boc-protected derivative of
the title compound (CHCl.sub.3 :METHANOL=80:1.fwdarw.50:1).
Yield: 5.20 g.
.sup.1 H-NMR (in CD.sub.30 D, 5:2 isomeric mixture): .delta.8.56, 8.31 &
8.18 (each 1H, each d), 7.97 & 7.87 (0.29 & 0.71H, each s), 7.58 (2H, q),
7.27 (1H, d), 4.14 (2H, t), 3.49 (2H, broad), 3.19 (2H, t), 3.10 (2H, t),
2.95-2.88 (4H, m), 2.88 (6H, s), 2.67 (2H, t), 2.00 & 1.95 (2.14 & 0.86H,
each s), 1.61 (2H, m), 1.41 (9H, s), 1.38-1.24 (18H, m), 0.90 (3H, t).
The Boc-protected derivative (5.25 g) was ice-cooled, added with
trifluoroacetic acid (30 ml) and dissolved under ice-cooling. And then the
mixture was left standing at room temperature for 0.5 hour. The
trifluoroacetic acid solution was slowly added dropwise to an aqueous
sodium hydrogen carbonate suspension (150 ml). A yellow fluorescent
viscous product was then deposited. The mixed solution was partitioned
with chloroform (100 ml). After the chloroform layer was washed with water
and dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=50:1.fwdarw.20:1) to give 3.56 g of a
residue. After the residue was dissolved in ethanol (150 ml) and added
with 1N hydrochloric acid (4.9 ml), the mixture was concentrated. The
residue was added with ethanol and concentrated again. The residue was
dissolved in chloroform (10 ml) and added dropwise to ether (150 ml). A
pale yellow amorphous deposit was deposited. After the ethereal layer was
discarded, the deposit was solidified with hexane. The powdery solid
obtained was collected by filtration and dried under reduced pressure to
give the title compound.
Yield: 3.30 g.
.sup.1 H-NMR (in CD.sub.3 OD, 10:1 isomeric mixture): .delta.8.57, 8.36 &
8.19 (each 1H, each d), 8.06 & 7.96 (0.1 & 0.9H, each s), 7.60 (2H, q),
7.34 (1H, d), 4.19 & 4.15 (1.8 & 0.2H, each t), 3.76 & 3.71 (0.2 & 1.8H,
each t), 3.17 (2H, t), 3.12 (2H, t), 2.97 (2H, t), 2.93 (6H, s), 2.91 (2H,
t), 2.72 (2H, m), 2.07 & 2.00 (2.7 & 0.3H, each s), 1.63 (2H, m), 1.39
(2H, m), 1.38-1.26 (18H, m), 0.89 (3H, t).
Example 20
(a)
N-[2-(dimethylamino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-methylbutyl)dithio]
-1-butenyl]formamide
In a solution of sodium hydroxide (1.0 g) in water (50 ml) was dissolved
Intermediate 9 (2.4 g), and the resulting solution was saturated with
sodium chloride. Sodium 1-methyl-butylthiosulfate (ca. 8 g) in the form of
powder was added to the aqueous solution. With stirring at room
temperature for about 15 minutes, no oily products were deposited. After
the reaction was partitioned with chloroform, the chloroform layer was
dried with anhydrous sodium sulfate and evaporated under reduced pressure
to give the title compound.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.03 & 8.02 (0.2
& 0.8H, each s), 3.79 (2H, t), 3.6-3.4 (2H, broad), 2.87 (2H, t), 2.78
(1H, m), 2.53 (2H, broad), 2.23 & 2.20 (4.8 & 1.2H, each s), 2.04 & 2.02
(2.4 & 0.6H, each s), 1.59 (1H, m), 1.48-1.36 (3H, m), 1.27 (3H, d), 0.91
(3H, m).
(b)
N-[2-(dimethylamino)ethyl]-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino
]propionyloxy]-1-methyl-2-[(1-methylbutyl)dithio]-1-butenyl]formamide
hydrochloride
In a solution of the residue obtained in the above step (a), Compound C
(2.0 g) and dimethylaminopyridine (300 mg) in tetrahydrofuran (30 ml) was
dissolved dicyclohexylcarbodiimide (1.34 g), and the mixture was stirred
at room temperature for about 3 hours. After white deposits were removed
by filtration, the filtrate was concentrated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :MeOH=40:1.fwdarw.20:1) to give 2.95 g of a fluorescent yellow
amorphous product. After the product was dissolved in ethanol and added
with 1N hydrochloric acid (4.6 ml), the solvent was evaporated under
reduced pressure. This procedure was repeated twice. The residue was next
dissolved in chloroform and concentrated again to give the title compound
as a fluorescent solid.
Yield: 2.70 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 6:1 isomeric mixture): .delta.8.62 (1H, d),
8.47, 8.28 & 8.11 (each 1H, each d), 8.06 & 7.85 (0.14 & 0.86H, each s),
7.61 (2H, q), 7.27 (1H, d), 3.9-3.8 (3H, m), 3.72 (2H, broad t), 3.15 (2H,
broad t), 2.84 (6H, s), 2.81 & 2.80 (5.14 & 0.86H, each s), 2.79 (1H, m),
2.65 (2H, broad t), 1.94 & 1.83 (2.57 & 0.43H, each s), 1.49 (1H, m),
1.44-1.3 (3H, m), 1.20 (3H, d), 1.13 (3H, d), 0.87 (3H, t).
Example 21
(a)
N-[2-(1-pyrrolidino)ethyl]-N-[4-hydroxy-1-methyl-2-[(1-methylbutyl)dithio]
-1-butenyl]formamide
In a solution of sodium hydroxide (1.32 g) in water (50 ml) was dissolved
Intermediate 10 (3.2 g), and the resulting solution was saturated with
sodium chloride. Sodium 1-methyl-butylthiosulfate (ca. 8 g) in the form of
powder was added to the aqueous solution. On stirring the mixture at room
temperature, oily products were deposited. After 15 minutes, the reaction
was partitioned with chloroform. After the chloroform layer was washed
with water and dried with anhydrous sodium sulfate, the solvent was
evaporated under reduced pressure to give the title compound.
(b)
N-[2-(1-pyrrolidino)ethyl]-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino
]propionyloxy]-1-methyl-2-[(1-methylbutyl)dithio]-1-butenyl]formamide
hydrochloride
The amorphous residue (3.4 g) obtained in the above step (a), Compound C
(3.03 g) and dimethylaminopyridine (300 mg) were dissolved in
tetrahydrofuran (30 ml). In this solution was dissolved
dicyclohexylcarbodiimide (1.98 g), and the mixture was stirred at room
temperature for about 3 hours. After white deposits were removed by
filtration, the filtrate was concentrated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=30:1.fwdarw.20:1) to give 3.10 g of a fluorescent
yellow amorphous product. After the product was dissolved in ethanol and
added with 1N hydrochloric acid (4.7 ml), the solvent was evaporated under
reduced pressure. This procedure was repeated twice. The residue obtained
was next dissolved in chloroform and concentrated again to give a
fluorescent solid as the title compound.
Yield: 2.95 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 5:1 isomeric mixture): .delta.8.62 (1H, d),
8.48, 8.30 & 8.11 (each 1H, each d), 8.07 & 7.84 (0.17 & 0.83H, each s),
7.61 (2H, q), 7.28 (1H, d), 3.9-3.8 (3H, m), 3.72 (2H, broad t), 3.56 (2H,
broad), 3.22 (2H, broad), 3.00 (2H, broad), 2.84 (7H, m), 2.64 (2H,
broad), 1.99 (2H, broad), 1.95 (3H, s), 1.85 (2H, m), 1.49 (1H, m),
1.44-1.3 (3H, m), 1.19 (3H, d), 1.14 (3H, d), 0.87 (3H, t).
Example 22
N,N'-{dithiobis[2-[(2-hydroxy)ethyl
]-1-methy-2,1-ethenediyl]}bis{N-[2-(tertbutyloxycarbonylaminoethyl]formami
de
An aqueous iodine-potassium iodide solution was prepared by adding iodine
(1.3 g) to a solution of potassium iodide (3.4 g) in distilled water (100
ml). At the same time, Intermediate 8 (3.7 g) was dissolved in distilled
water (350 ml) and added with 1N hydrochloric acid (21 ml). To this
solution was slowly added dropwise the aqueous iodine-potassium iodide
solution at room temperature. After the addition of the aqueous
iodine-potassium iodide solution was completed, the mixture was stirred
for 15 minutes and then partitioned with ethyl acetate (50 ml). The ethyl
acetate layer was further washed with an aqueous potassium iodide solution
and dried over anhydrous sodium sulfate. And then the solvent was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography to give the title compound as a colorless
solid (1.95 g).
.sup.1 H-NMR (in CD.sub.3 OD): .delta.8.02 & 7.92 (1H, s), 3.72 (2H, m),
3.51 (2H, broad m), 3.20 (2H, broad t), 2.78 (2H, broad t), 2.07-2.03 (3H,
m), 1.43 (9H, s).
Example 23
N[(2-hydroxy)ethyl]-N-[2-[(2-propyl)dithio]ethenyl]formamide
To a solution of sodium hydroxide (3.2 g) in distilled water (50 ml) was
added Intermediate 2 (8.2 g), and the mixture was left standing at room
temperature for 10 minutes. Sodium 2-propylthiosulfate (22 g) in the form
of powder was added to the aqueous solution. Immediately after the
addition, a pale yellow oily product was deposited. The reaction was
partitioned with chloroform (100 ml). After the chloroform layer was dried
with anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography to give 75 g of the title compound.
.sup.1 H-NMR (in CDCl.sub.3): .delta.8.33 (1H, s), 6.14 (1H, d), 5.91 (1H,
d), 3.75 (2H, t), 3.09 (1H, m), 1.33 (3H, s), 1.32 (3H, s).
Example 24
(a) N-(2-hydroxyethyl)-N-[2-(isobutyrylthio)vinyl]formamide
In a solution of sodium hydroxide (1.32 g) in water (30 ml) was dissolved
Intermediate 2 (3.2 g). To the mixed solution ice-cooled, dioxane (20 ml)
was added. And then the mixed solution of isobutyryl chloride (1.60 g) and
dioxane (10 ml) was added in one portion. The reaction mixture immediately
after the addition had a pH of about 7. The reaction was extracted with
chloroform (200 ml). After the chloroform layer was washed twice with
water and dried with anhydrous sodium sulfate, the solvent was evaporated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1) to give the
title compound.
Yield: 2.59 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.34 & 8.12 (0.8
& 0.2H, each s), 6.81, 6.42, 6.33 & 6.30 (0.2, 0.8, 0.8 & 0.2H, each d,
J=9, 8.5, 8.5 & 9 Hz), 3.79 (2H, t), 3.75 (2H, m), 2.78 (1H, m), 2.13 &
1.75 (0.8 & 0.2H, each broad t), 1.24 & 1.23 (6H, each d).
(b) N
[2-[[3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyloxy]ethyl]-N-[2-(i
sobutyrylthio)vinyl]formamide
The compound (2.59 g) obtained in the above step (a), Compound B (3.23 g)
and dimethylaminopyridine (250 mg) were dissolved in tetrahydrofuran (30
ml). In this solution was dissolved dicyclohexylcarbodiimide (2.3 g), and
the mixture was left standing at room temperature for about 13 hours.
After white deposits were removed by filtration, the filtrate was
concentrated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :acetone=10:1) to give the
title compound.
Yield: 1.72 g.
.sup.1 H-NMR (in CDCl.sub.3, 5:1 isomeric mixture): .delta.8.55 & 8.30
(each 1H, each d), 8.25 (1H, s), 8.24 (1H, m), 7.60-7.51 (2H, m), 7.19
(1H, d), 6.72 & 6.30 (0.17 & 1.83H, d & m), 5.58 & 5.29 (0.87 & 0.17H,
broad t), 4.13 & 4.09 (1.83 & 0.17H, each t), 3.78 (2H, t), 3.15 (2H, q),
2.89 (6H, s), 2.77 (1H, m), 2.44 (2H, t.), 1.23 & 1.22 (6H, each d).
Example 25
(a) N-(2-hydroxyethyl)-N-[2-(pivaloylthio)vinyl]formamide
In a solution of sodium hydroxide (1.32 g) in water (30 ml) was dissolved
Intermediate 2 (3.2 g). To the mixed solution ice-cooled, dioxane (20 ml)
was added. And then the mixed solution of pivaloyl chloride (1.81 g) and
dioxane (10 ml) was added in one portion. The reaction mixture immediately
after the addition had a pH of about 7. The reaction was extracted with
chloroform (150 ml). After the chloroform layer was washed twice with
water and dried with anhydrous sodium sulfate, the solvent was evaporated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1) to give the
title compound.
Yield: 3.04 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.36 & 8.12 (0.8
& 0.2H, each s), 6.82, 6.42, 6.34 & 6.31 (0.2, 0.8, 0.8 & 0.2H, each d,
J=9, 8.5, 8.5 & 9 Hz), 3.79 (2H, q), 3.75 (2H, m), 2.13 & 1.71 (0.8 &
0.2H, each broad t), 1.27 (9H, s).
.sup.1 H-NMR (in DMSO-d.sub.6, 6:5 isomeric mixture): .delta.8.33 & 8.07
(0.55 & 0.45H, each s, --N--CHO), 6.88, 6.77, 6.03 & 5.97 (0.45, 0.55,
0.45 & 0.55H, each d, J=9, 8.5, 9 & 8.5 Hz, --CH.dbd.CH--), 3.67 & 3.60
(each 1H, each t, --O--CH.sub.2 CH2--), 3.45 (2H, m, --O--CH.sub.2
CH.sub.2 --), 1.22 (9H, s, --C(CH.sub.3).sub.3).
(b)
N-[2-[[3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyl]oxy]ethyl]-N-[2
-(pivaloylthio)vinyl]formamide
The compound (2.31 g) obtained in the above step (a), Compound B (2.65 g)
and dimethylaminopyridine (250 mg) were dissolved in tetrahydrofuran (30
ml). In this solution was dissolved dicyclohexylcarbodiimide (2.0 g), and
the mixture was left standing at room temperature for about 12 hours.
After white deposits were removed by filtration, the filtrate was
concentrated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :acetone=10:1) to give the
title compound.
Yield: 2.11 g.
.sup.1 H-NMR (in CDCl.sub.3, 5:1 isomeric mixture): .delta.8.55 & 8.30
(each 1H, each d), 8.26 (1H, s), 8.24 (1H, m), 7.60-7.51 (2H, m), 7.19
(1H, d), 6.74 & 6.31 (0.4 & 1.6H, d & m), 5.58 & 5.28 (0.8 & 0.2H, broad
t), 4.13 & 4.09 (1.6 & 0.4H, each t), 3.78 (2H, t), 3.14 (2H, q), 2.89
(6H, s), 2.46 & 2.43 (0.4 & 1.6H, each t), 1.26 & 1.25 (9H, each s).
Example 26
(a) N-(2-hydroxyethyl)-N-[2-(benzoylthio)vinyl]formamide
In a solution of sodium hydroxide (1.76 g) in water (30 ml) was dissolved
Intermediate 2 (4.2 g). To the mixed solution ice-cooled, dioxane (20 ml)
was added. And then the mixed solution of benzoyl chloride (2.81 g) and
dioxane (10 ml) was added in one portion. The reaction mixture immediately
after the addition had a pH of about 7. The reaction was extracted with
chloroform. After the-chloroform layer was washed twice with water and
dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1) to give the title
compound.
Yield: 4.14 g.
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.43 & 8.16
(0.75 & 0.25H, each s), 7.77, 7.63 & 7.50 (2, 1 & 2H, each m), 6.92, 6.58,
6.54 & 6.52 (0.25, 0.75, 0.25 & 0.75H, each d, J=9, 8.5, 8.5 & 9 Hz), 3.81
(4H, m), 2.18 & 1.87 (0.75 & 0.25H, each broad t).
.sup.1 H-NMR (in DMSO-d.sub.6, 10:9 isomeric mixture): .delta.8.43 & 8.12
(0.53 & 0.47H, each s), 7.97, 7.74 & 7.60 (2, 1 & 2H, each m), 7.00, 6.91,
6.29 & 6.18 (0.47, 0.53, 0.47 & 0.53H, each d, each J=9 Hz), 4.93 & 4.86
(0.47 & 0.53H, each broad t), 3.74 & 3.69 (each 1H, each t), 3.52 & 3.48
(each 1H, each q).
(b)
N-[2-[[3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyl]oxy]ethyl]-N-[2
-(benzoylthio)vinyl]formamide
The compound (2.5 g) obtained in the above step (a), Compound B (2.92 g)
and dimethylaminopyridine (250 mg) were dissolved in tetrahydrofuran (20
ml). In this solution was dissolved dicyclohexylcarbodiimide (2.2 g), and
the mixture was left standing at room temperature for about 14 hours.
After white deposits were removed by filtration, the filtrate was
concentrated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :acetone=50:3.fwdarw.10:1) to
give the title compound.
Yield: 3.70 g.
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.54 (1H, d),
8.34 & 7.99 (0.75 & 0.25H, each s), 8.29 (1H, d), 8.23 (1H, dd), 7.95 (2H,
m), 7.65-7.47 (5H, m), 7.18 (1H, d), 6.83, 6.55, 6.53 & 6.39 (0.25, 0.75,
0.25 & 0.75H, each d, J=9, 8, 9 & 8 Hz), 5.57 & 5.27 (0.75 & 0.25H, broad
t), 4.16 & 4.12 (1.5 & 0.5H, each t), 3.83 (2H, t), 3.14 (2H, q), 2.89
(6H, s), 2.46 & 2.43 (0.5 & 1.5H, each t).
Example 27
(a) N-(2-hydroxy-1-propyl)-N-[1-methyl-2-(pivaloylthio)vinyl]formamide
In a solution of sodium hydroxide (1.6 g) in water (30 ml) was dissolved
Intermediate 11 (4.3 g). To the mixed solution ice-cooled, dioxane (20 ml)
was added. And then the mixed solution of pivaloyl chloride (2.0 g) and
dioxane (10 ml) was added in one portion (the reaction mixture immediately
after the addition had a pH of 7). After the reaction was extracted with
chloroform. The chloroform layer was washed with water and dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=50:1.fwdarw.40:1) to give the title
compound.
Yield: 2.47 g.
.sup.1 H-NMR (in CDCl.sub.3, 7:1 isomeric mixture): .delta.8.10 & 8.04
(0.87 & 0.13H, each s), 6.45 (1H, d, J=1 Hz), 3.98 (1H, broad), 3.54, 3.42
& 3.31 (0.87, 0.87 & 0.26H, dd, dd & m), 2.66 (1H, broad), 2.08 & 2.04
(2.62 & 0.38H, d, J=1.5 Hz), 1.24 (9H, s), 1.21 (3H, d).
(b)
N-[2-[[3-[(5-dimethylaminonaphthylsulfonyl)amino]propionyl]oxy]1-propyl]-N
-[1-methyl-2-(pivaloylthio)vinyl]formamide
Compound B (2.6 g), the compound (2.1 g) obtained in the above step (a) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
In this solution was dissolved dicyclohexylcarbodiimide (1.68 g), and the
mixture was left standing at room temperature for about 12 hours. After
white deposits were removed by filtration, the filtrate was concentrated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography (CHCl.sub.3 :acetone=10:1) to give the title
compound.
Yield: 3.51 g.
.sup.1 H-NMR (in CDCl.sub.3, 4:1 isomeric mixture): .delta.8.55, 8.33 &
8.29 (each 1H, each d), 8.04 & 7.93 (0.8 & 0.2H, each s), 7.58 & 7.53
(each 1H, each 7.20 (1H, d), 6.63 & 6.38 (0.2 & 0.8H, each s), 5.71 & 5.65
(0.8 & 0.2H, each broad t), 4.91 & 4.84 (0.8 & 0.2H, each m), 3.67 & 3.52
(each 1H, each dd), 3.12 (2H, m), 2.90 (6H, s), 2.46 & 2.41 (0.4 & 1.6H,
each m), 2.03 & 2.02 (0.6 & 2.4H, each s), 1.23, 1.19 & 1.17 (12H, s, s &
d).
Example 28
(a)
N-[2-(dimethylamino)ethyl]-N-[4-hydroxy-1-methyl-2-(pivaloylthio)-1-buteny
l]formamide
In a solution of sodium hydroxide (1.86 g) in water (20 ml) was dissolved
Intermediate 9 (4.31 g). To the mixed solution ice-cooled, dioxane (20 ml)
was added. And then the mixed solution of pivaloyl chloride (1.80 g) and
dioxane (10 ml) was added in one portion (the reaction mixture immediately
after the addition had a pH of 9). The reaction was partitioned with
chloroform (150 ml). After the chloroform layer was washed with water and
dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure to give the title compound.
(b)
N-[2-(dimethylamino)ethyl]-N-[4-[2-[(5-dimethylaminonaphthylsulfonyl)amino
]propionyl]-1-methyl-2-(pivaloylthio)-1-butenyl]formamide hydrochloride
The compound (2.3 g) obtained in the above step (a), Compound C (2.3 g) and
dimethylaminopyridine (300 mg) were dissolved in tetrahydrofuran (30 ml).
In this solution was dissolved dicyclohexylcarbodiimide (1.50 g), and the
mixture was stirred at room temperature for about 3 hours. After white
deposits were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:2.fwdarw.10:3). The purified
product (1.24 g) was dissolved in ethanol and added with 1N hydrochloric
acid (2.0 ml), and the mixture was concentrated udder reduced pressure.
This procedure was repeated twice. The residue was next dissolved in
chloroform and concentrated under reduced pressure again to give a
fluorescent amorphous solid as the title compound.
Yield: 1.10 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.10.12 (1H,
broad), 8.60 (1H, d), 8.48 & 8.28 (each 1H, each d), 8.10 (1H, dd), 8.02 &
7.67 (0.1 & 0.9H, s), 7.60 (2H, q), 7.28 (1H, d), 3.89-3.78 (3H, m), 3.68
(2H, broad t), 2.99 (2H, broad), 2.84 (6H, s), 2.78 & 2.77 (each 3H, each
s), 2.42 (2H, broad t), 2.04 & 2.00 (2.7 & 0.3H, each s), 1.17 (9H, s),
1.12 (3H, d).
Example 29
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(tetrahy
drofurfuryl)dithio]-1-butenyl]formamide
This compound was synthesized in accordance with the report (1) by
Matsukawa et al. (supra). In brief, sodium tetrahydrofurfurylthiosulfate
(22 g) in the form of powder was added to an aqueous yellow solution
obtained by adding a suspension of thiamine hydrochloride (23.6 g) in
water (20 ml) to an aqueous solution (20 ml) of sodium hydroxide (8.4 g).
Pale yellow amorphous product thus deposited was washed with water,
dissolved in ethyl acetate and partitioned with a saturated aqueous sodium
sulfate solution containing sodium hydrogen carbonate. The ethyl acetate
layer was concentrated under reduced pressure, And then white solids
deposited were collected by filtration and dried under reduced pressure to
give the title compound.
.sup.1 H-NMR (in CDCl.sub.3): .delta.8.0 (1H, s), 7.8 (1H, s), 3.88 (1H,
m), 3.81 (1H, m), 3.70 (1H, m), 3.63 (2H, t), 2.85 (2H, m), 2.57 (2H, d),
2.39 (3H, s), 2.09 (3H, s), 2.01 (1H, m), 1.90 (2H, m), 1.56 (1H, m).
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-amino-3-(3,4-dipivaloylox
yphenyl)propionyl]oxy-1-methyl-2-[(tetrahydrofurfuryl)dithio]-1-butenyl]for
mamide
Compound D (3.5 g), the compound (3.0 g) obtained in the above step (a) and
dimethylaminopyridine (97 mg) were dissolved in a mixture of
acetonitrile:tetrahydrofuran=30:20 ml. In this solution was dissolved
dicyclohexylcarbodiimide (1.50 g) under ice-cooling, and the mixture was
stirred for about 3 hours. After white deposits were removed by
filtration, the filtrate was concentrated under reduced pressure. Pale
yellow amorphous product as the residue obtained was purified by silica
gel column chromatography (CHCl.sub.3 :methanol=40:1) to give the
Boc-protected derivative of the title compound.
Yield: 4.07 g.
The Boc-protected derivative (3.80 g) was added with trifluoroacetic acid
(30 ml) and dissolved under ice-cooling. After the derivative was
dissolved, the solution was left standing at room temperature for about 15
minutes and added dropwise to a saturated suspension of sodium hydrogen
carbonate prepared separately (250 ml). After the mixture was partitioned
with ethyl acetate (ca. 100 ml), the ethyl acetate layer was washed with
an aqueous sodium sulfate solution and dried over anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure to give a white
solid residue (3.13 g). After the residue was dissolved in ethanol (ca. 50
ml) and added with 1N hydrochloric acid (4.2 ml), the solvent was removed
under reduced pressure. To the amorphous product thus obtained was added
ethanol (100 ml) again, and the solvent was evaporated again under reduced
pressure to give amorphous product. Ether and hexane were added to the
residue, and white powder obtained was collected by filtration and dried
under reduced pressure to give the title compound.
Yield: 2.94 g.
.sup.1 H-NMR (in CD.sub.3 OD): .delta.7.96 (1H, s), 7.85 (1H, s), 7.23-7.13
(3H, m), 4.33 (1H, m), 4.29 (1H, m), 4.15 (1H, m), 3.88 (1H, m), 3.81 (1H,
m), 3.70 (1H, m), 3.21 (2H, m), 2.93 (2H, m), 2.63 (2H, m), 2.41 (3H, s),
2.04 (3H, s), 2.00 (1H, m), 1.90 (2H, m), 1.57 (1H, m), 1.34 & 1.33 (18H,
each s).
Example 30
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(2-propy
l)dithio]-1-butenyl]formamide
Sodium isopropylthiosulfate (22 g) in the form of powder was added to an
aqueous yellow solution obtained by adding a suspension of thiamine
hydrochloride (23.6 g) in water (20 ml) to an aqueous solution (20 ml) of
sodium hydroxide (8.4 g). Pale yellow amorphous product deposited was
washed with water, dissolved in ethyl acetate and partitioned with a
saturated aqueous sodium sulfate solution containing sodium hydrogen
carbonate. The ethyl acetate layer was concentrated under reduced
pressure, and white solids deposited were collected by filtration and
dried under reduced pressure to give 12.5 g of the title compound.
.sup.1 H-NMR (in CDCl.sub.3): .delta.8.02 (1H, s), 7.84 (1H, s), 3.61 (2H,
t), 2.84 (2H, t), 2.61 (1H, m), 2.38 (3H, s), 2.10 (3H, s), 1.10 (6H, d).
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-amino-3-(3,4-dipivaloylox
yphenyl)propionyl]oxy-1-methyl-2-[(2-propyl)dithio]-1-butenyl]formamide
hydrochloride
The compound (2.3 g) obtained in the above step (a), Compound D (2.8 g),
and dimethylaminopyridine (100 mg) were dissolved in acetonitrile and
ice-cooled. In this solution was dissolved dicyclohexylcarbodiimide (1.30
g), and the mixture was left standing under ice-cooling for 3 hours. After
white deposits were removed by filtration, the filtrate was concentrated
under reduced pressure. The residue obtained was purified by silica gel
column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1) to give the
Boc-protected derivative of the title compound.
Yield: 4.50 g.
.sup.1 H-NMR (in CDCl.sub.3): .delta.7.93 & 7.81 (each 1H, each s),
7.06-6.92 (3H, m), 4.52 (1H, broad d), 4.15 (2H, broad d), 3.16 (2H, broad
t), 2.88 (2H, broad), 2.59 (1H, m), 2.44 (3H, s), 1.96 (3H, s), 1.42 (6H,
s), 1.33 (9H, s), 1.10 (6H,d).
The Boc-protected derivative (3.61 g) was dissolved in trifluoroacetic acid
(20 ml) under ice-cooling. After the solution was left standing at room
temperature for about 15 minutes, it was added to a sodium hydrogen
carbonate suspension (200 ml). White amorphous product was deposited. The
mixture was partitioned with ethyl acetate. The aqueous layer was
partitioned with ethyl acetate. The combined ethyl acetate layers were
further partitioned with 0.5N hydrochloric acid. The aqueous layer
obtained was adjusted to a pH of 8-9 with aqueous sodium hydrogen
carbonate and partitioned with ethyl acetate. The ethyl acetate layer was
dried over anhydrous sodium sulfate and evaporated under reduced pressure
(3.04 g). The residue was dissolved in ethanol (150 ml) and added with 1N
hydrochloric acid (4.3 ml), and the solvent was removed under reduced
pressure. To the residue obtained was added the same amount of ethanol,
and the mixture was concentrated again to amorphous product. To the
product was added a small amount of ether, and the mixture was added
dropwise to hexane (200 ml). White powder deposited was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 3.09 g.
.sup.1 H-NMR (in CD.sub.3 OD): .delta.7.99 & 7.87 (each 1H, each s),
7.23-7.13 (3H, m), 4.3 (1H, t), 4.3 & 4.2 (each 1H, each m), 3.22 & 3.21
(each 1H, each d), 2.93 (1H, m), 2.70 (1H, m), 2.41 (3H, s), 2.05 (3H, s),
1.34 (18H, s), 1.12 (6H, d)
(c)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
)amino]ethylaminocarbonyl]oxy-1-methyl-2-[(2-propyl)dithio]-1-butenyl]forma
mide
In a solution of the compound obtained in the above step (a) (3.57 g) in
tetrahydrofuran (30 ml) was dissolved carbonyldiimidazole (1.63 g) in the
form of powder, and the mixture was left standing at room temperature for
about 15 minutes. To this solution was added Compound A (2.95 g) in the
form of powder, and the mixture was stirred at room temperature for 13
hours. After insolubles were removed by filtration, and the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=80:1.fwdarw.30:1)
to give the title compound.
Yield: 6.17 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.8.46 & 8.26 (each 1H, each d), 8.12
(1H, broad), 8.09 (1H, dd), 7.91 & 7.85 (each 1H, each s), 7.62 & 7.58
(each 1H, each dd), 7.25 (1H, d), 7.09 (1H, broad t), 6.79 (2H, broad),
4.37 (2H, broad), 3.91 (2H, t), 3.00 (2H, q), 2.83 (8H, m), 2.78 (2H, t),
2.61 (1H, m), 2.24 & 1.99 (each 3H, each s), 1.02 (6H, d, J=6.5 Hz).
Example 31
(a)
N-methyl-N-[[4-[2-amino-3-(3,4-di-O-pivaloylphenyl)propionyl]oxy-1-methyl-
2-[(1-propyl)dithio]-1-butenyl]formamidehydrochloride
This compound was synthesized by the same method as described in Example
29.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
)amino]ethylaminocarbonyl
]oxy-1-methyl-2-[(1-propyl)dithio]-1-butenyl]formamide
In a solution of the compound obtained in the above step (a) (3.57 g) in
tetrahydrofuran (20 ml) was dissolved carbonyldiimidazole (1.63 g) in the
form of powder, and the mixture was left standing at room temperature for
about 15 minutes. To this solution was added Compound A (2.95 g) in the
form of powder, and the mixture was stirred at room temperature for 15
hours. After insolubles were removed by filtration, the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1)
to give the title compound.
Yield: 5.73 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.8.46 & 8.26 (each 1H, each d), 8.14
(1H, broad), 8.09 (1H, dd), 7.88 & 7.84 (each 1H, each s), 7.62 & 7.58
(each 1H, each dd), 7.25 (1H, d), 7.10 (1H, broad t), 6.77 (2H, broad),
4.37 (2H, broad), 3.93 (2H, t), 3.00 (2H, q), 2.83 (8H, m), 2.77 (2H, t),
2.33 (2H, t), 2.24 & 1.99 (each 3H, each s), 1.40 (2H, m), 0.84 (3H, t).
Example 32
(a)
N-methyl-N-[[4-[2-amino-3-(3,4-di-O-pivaloylphenyl)propionyl]oxy-1-methyl-
2-[(1-methylbutyl)dithio]-1-butenyl]formamide hydrochloride
This compound was synthesized by the same method as described in Example
29.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
)amino]ethylaminocarbonyl]oxy-1-methyl-2-[(1-methylbutyl)dithio]-1-butenyl]
formamide hydrochloride
In a suspension of the compound obtained in the above step (a) (2.5 g) in
tetrahydrofuran (15 ml) was dissolved carbonyldiimidazole (1.17 g), and
the mixture was left standing at room temperature for 1 hour. To this
solution was added Compound A (2.2 g), and the mixture was stirred at room
temperature for about 16 hours. After insolubles and deposits were removed
by filtration, the filtrate was evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=60:1.fwdarw.40:1). The residue (4.37 g) obtained was
dissolved in chloroform and added with a solution of 4N hydrochloric acid
in dioxane (1.6 ml). The mixture was concentrated under reduced pressure.
The residue was dissolved in chloroform and added dropwise to ether. White
to greenish white fluorescent powder obtained was collected by filtration
and dried under reduced pressure to give the title compound.
Yield: 3.93 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.29 (1H, broad), 8.48 & 8.29 (each
1H, each d), 8.25 (2H, broad), 8.23 (1H, s), 8.09 (1H, dd), 8.03 (1H,
broad t), 7.97 (1H, s), 7.62 (2H, m), 7.29 (1H, d), 7.16 (1H, broad t),
4.48 (2H, broad), 3.96 (2H, t), 3.01 (2H, q), 2.85 (6H, s), 2.81 (4H, m),
2.65 (1H, m), 2.02 (3H, s), 1.36-1.24 (4H, m), 1.05 (3H, d), 0.83 (3H, t).
(c)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(1-methylbutyl)dithio]-1-butenyl]f
ormamide hydrochloride
The solution of the sodium salt of Compound B (4.0 g) in water (40 ml) was
saturated with sodium sulfate. This aqueous solution was adjusted to a pH
of ca. 3 with 4N hydrochloric acid, partitioned with chloroform to extract
a fluorescent substance. After the chloroform layer was dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The fluorescent residue obtained (2.73 g), dimethylaminopyridine
(250 mg) and the compound prepared in the above step (a) (3.26 g) were
dissolved in the mixture of tetrahydrofuran (15 ml) and acetonitrile (10
ml). Dicyclohexylcarbodiimide (1.86 g) was added, and the resulting
solution was left standing at room temperature for about 15 hours. After
white insolubles were removed by filtration, the filtrate was evaporated
under reduced pressure. The residue obtained was dissolved in chloroform
and purified by silica gel column chromatography (CHCl.sub.3
:methanol=60:1.fwdarw.40:1). The purified product (4.00 g) obtained was
dissolved in chloroform and added with a solution of 4N hydrochloric acid
in dioxane (1.45 ml), and the mixture was concentrated under reduced
pressure. The residue obtained was dissolved in chloroform and added
dropwise to ether. Pale green powdery crystals were collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 3.91 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.9.30 (1H,
broad), 8.48 (1H, d), 8.27 (3H, broad), 8.22 (1H, s), 8.11 (1H, dd), 8.06
(1H, broad t), 7.96 (1H, s), 7.63 & 7.59 (each 1H, each dd), 7.28 (1H, d),
4.47 (2H, broad), 3.98 (2H, t), 3.02 (2H, q), 2.84 (6H, s), 2.80 (2H, t),
2.64 (1H, m), 2.44 (2H, t), 2.02 & 1.82 (2.7 & 0.3H, each s), 1.36-1.24
(4H, m), 1.05 (3H, d), 0.83 (3H, t).
(d)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(1-methylbutyl)dithio]-1-butenyl]f
ormamide hydrochloride
The compound prepared in the above step (a) (2.0 g), Compound C (1.8 g) and
dimethylaminopyridine (250 mg) were suspended in the mixture of
tetrahydrofuran (20 ml) and dichloromethane (20 ml). In this suspension
was dissolved dicyclohexylcarbodiimide (1.24 g), and the mixture was left
standing at room temperature for about hours. After white insolubles were
removed by filtration, the filtrate was concentrated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (developed with CHCl.sub.3 :acetone=10:1, then eluted with
CHCl.sub.3 :methanol=45:1.fwdarw.35:1) to give a purified product (3.53
g). The product was dissolved in ethanol and added with 1N hydrochloric
acid, and the mixture was concentrated under reduced pressure. The residue
was taken in ethanol and concentrated again to give a yellowish white
amorphous product. The product was dissolved in chloroform and added
dropwise to ether. Greenish white powder crystallized was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 2.25 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 10: 1 isomeric mixture): .delta.9.29 (1H,
broad), 8.63 (1H, d), 8.48 (1H, d), 8.29 (3H, broad d), 8.23 (1H, s), 8.11
(1H, dd), 7.95 (1H, s), 7.61 (2H, q), 7.28 (1H, d), 4.47 (2H, broad), 3.86
(2H, m), 3.77 (1H, m), 2.84 (6H, s), 2.62 (4H, m), 2.00 & 1.79 (2.7 &
0.3H, each s), 1.34-1.24 (4H, m), 1.13 (3H, d), 1.04 (3H, d), 0.84 (3H,
t).
Example 33
(a)
N-methyl-N-[[4-hydroxy-1-methyl-2-[1-(ethyl)propyldithio]-1-butenyl]formam
ide hydrochloride
This compound was synthesized in the same manner as described in Example
29.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
)amino]ethylaminocarbonyloxy-1-methyl-2-[(1-ethylpropyl)dithio]-1butenyl]fo
rmamide hydrochloride
In a suspension of the compound obtained in the above step (a) (2.0 g) in
tetrahydrofuran (15 ml) was dissolved carbonyldiimidazole (0.90 g). After
0.5 hour, complete consumption of the compound of the above step (a) was
confirmed by TLC. Compound A (1.76 g) was added to the reaction. The
mixture was stirred at room temperature for about 13 hours. After
insolubles were removed by filtration, the filtrate was concentrated under
reduced pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1). The purified
product (2.97 g) was dissolved in chloroform and added with a solution of
4N hydrochloric acid in dioxane (1.05 ml), and the mixture was
concentrated under reduced pressure. The residue obtained was dissolved in
chloroform and added dropwise to ether. White to greenish white
fluorescent powder obtained was collected by filtration and dried under
reduced pressure to give the title compound.
Yield: 2.12 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 10:1 isomeric mixture): .delta.9.31 (1H,
broad), 8.50 & 8.28 (each 1H, each d), 8.26 (2H, broad), 8.23 (1H, s),
8.10 (1H, dd), 8.05 (1H, broad t), 7.96 (1H, s), 7.63 (2H, m), 7.31 (1H,
broad d), 7.19 (1H, broad t), 4.47 (2H, broad), 3.97 (2H, t), 3.01 (3H,
q), 2.86 (6H, s), 2.81 (4H, m), 2.42 (1H, m), 2.02 & 1.82 (0.9 & 0.1H,
each s), 1.38 (4H, m), 0.83 (6H, t).
(c)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulphonyl)amino]propionyl]oxy-1-methyl-2-[(1-ethylpropyl)dithio]-1-butenyl]
formamide hydrochloride
The solution of the sodium salt of Compound B (3.0 g) in water (40 ml) was
saturated with sodium sulfate. This aqueous solution was adjusted to a pH
of ca. 3 with 4N hydrochloric acid, partitioned with chloroform to extract
a fluorescent substance. After the chloroform layer was dried with
anhydrous sodium sulfate, the solvent was evaporated under reduced
pressure. The residue obtained (1.67 g), dimethylaminopyridine (200 mg)
and the compound prepared in the above step (a) (2.0 g) were dissolved in
the mixture of tetrahydrofuran (15 ml) and acetonitrile (10 ml).
Dicyclohexylcarbodiimide (1.24 g) was added, and the resulting solution
was left standing at room temperature for about 12 hours. After white
insolubles were removed by filtration, the filtrate was evaporated under
reduced pressure. The residue obtained was dissolved in chloroform and
purified by silica gel column chromatography (CHCl.sub.3
:methanol=60:1.fwdarw.40:1). The purified product (2.14 g) was dissolved
in chloroform and added with a solution of 4N hydrochloric acid in dioxane
(0.77 ml). The mixture was concentrated under reduced pressure. The
residue obtained was dissolved in chloroform and added dropwise to ether.
Pale green powdery crystals were collected by filtration and dried under
reduced pressure to give the title compound.
Yield: 2.06 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.9.30 (1H,
broad), 8.49 (1H, d), 8.28 (3H, broad), 8.21 (1H, s), 8.11 (1H, dd), 8.06
(1H, broad t), 7.95 (1H, s), 7.63 & 7.59 (each 1H, t), 7.29 (1H, broad d),
4.47 (2H, broad), 4.00 (2H, t), 3.02 (2H, q), 2.85 (6H, s), 2.81 (2H, t),
2.44 (3H, m), 2.01 & 1.82 (0.9 & 0.1H, each s), 1.38 (4H, m), 0.84 (6H,
t).
Example 34
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(1-penty
l)dithio]-1-butenyl]formamide
This compound was synthesized in the same manner as described in Example
29.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(1-pentyl)dithio]-1-butenyl]formam
ide hydrochloride
A solution of the sodium salt of Compound B (4.0 g) in water (40 ml) was
saturated with sodium slufate. To this solution was added 4N hydrochloric
acid to adjust the pH to about 3, and the mixture was partitioned with
chloroform to extract a fluorescent substance. After the chloroform layer
was dried with anhydrous sodium sulfate, the solvent was evaporated under
reduced pressure. The fluorescent residue obtained (2.80 g),
dimethylaminopyridine (250 mg) and the compound obtained in the above step
(a) (3.34 g) were dissolved in the mixture of tetrahydrofuran (15 ml) and
acetonitrile (10 ml). After dicyclohexylcarbodiimide (1.86 g) was added,
the mixture was left standing at room temperature for about 12 hours.
White insolubles were removed by filtration, and the filtrate was
concentrated under reduced pressure. The residue was dissolved in
chloroform and purified by silica gel column chromatography (CHCl.sub.3
:methanol=60:1.fwdarw.40:1). The purified product obtained (4.19 g) was
dissolved in chloroform and added with a solution of 4N hydrochloric acid
in dioxane (1.5 ml). The mixture was concentrated under reduced pressure.
The residue was dissolved in chloroform and added dropwise to ether. Pale
green powdery crystals obtained was collected by filtration and dried
under reduced pressure to give the title compound.
Yield: 4.13 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.9.27 (1H,
broad), 8.49 & 8.28 (each 1H, each d), 8.24 (2H, broad), 8.20 (1H, s),
8.12 (1H, d), 8.07 (1H, broad), 7.94 (1H, s), 7.64 & 7.60 (each 1H, each
t), 7.29 (1H, d), 4.47 (2H, broad), 4.01 (2H, t), 3.03 (2H, q), 2.85 (6H,
s), 2.79 (2H, t), 2.45 (2H, t), 2.01 & 1.82 (2.7 & 0.3H, each s), 1.43
(2H, m), 1.24 (4H, m), 0.84 (3H, m).
Example 35
(a) N.[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-
hydroxy-1-methyl-2-[(2-propenyl)dithio]-1-butenyl]formamide
This compound was synthesized in the same manner as described in Example
29.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-amino-3-(3,4-dipivaloylox
yphenyl)propionyl]oxy-1-methyl-2-[(2-propenyl)dithio]-1-butenyl]formamide
This compound was synthesized with the compound prepared in the above step
(a) in the same manner as described in Example 30 (b).
.sup.1 H-NMR (in CDCl.sub.3): .delta.7.97 (1H, s), 7.86 (1H, s), 7.23-7.12
(3H, m), 5.72-5.64 (1H, m), 5.16-5.10 (2H, m), 4.34 (1H, t), 4.29 (1H, m),
4.16 (1H, m), 3.22 (2H, t), 3.11 (32H, d), 2.91 (2H, m), 2.40 (3H, s),
2.04 (3H, s), 1.34 (18H, s).
Example 36
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(1-butyr
yl)thio]-1-butenyl]formamide
This compound was synthesized in accordance with the report (2) by
Matsukawa et at. (supra).
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(1-butyryl)thio]-1-butenyl]formami
de
The sodium salt of Compound B (5.2 g) was dissolved in water (100 ml). To
this solution was added 4N hydrochloric acid to adjust the pH to 2-3, and
the yellowish green insolubles were extracted with chloroform. After the
chloroform layer was dried with anhydrous sodium sulfate, the solvent was
evaporated under reduced pressure. The residue obtained, the compound
obtained in the above step (a) (3.53 g) and dimethylaminopyridine (250 mg)
were dissolved in the mixed solvent of tetrahydrofuran (25 ml),
acetonitrile (10 ml) and dichloromethane (20 ml). To the mixed solution
was added dicyclohexylcarbodiimide (2.1 g), and the mixture was stirred at
room temperature for about 16 hours. After insolubles were removed by
filtration, the filtrate was concentrated under reduced pressure. The
residue was purified by silica gel column chromatography (CHCl.sub.3
:methanol=70:1.fwdarw.40:1) to give the title compound.
Yield: 2.86 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 9:1 isomeric mixture): .delta.8.47, 8.25 &
8.11 (each 1H, each d), 8.08 (1H, t), 7.79 & 7.74 (0.1 & 0.9H, each s),
7.73 & 7.71 (0.1 & 0.9H, each s), 7.63 & 7.58 (each 1H, each t), 7.25 (1H,
d), 6.68 (2H, broad), 4.36 (2H, broad), 3.88 (2H, t), 3.01 (2H, q), 2.83
(6H, s), 2.53 (2H, t), 2.42-2.36 (4H, m), 2.24 & 2.01 (each 3H, s), 1.45
(2H, m), 0.82 (3H, t).
Example 37
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(iso-but
yryl)thio]-1-butenyl]formamide
This compound was synthesized in the same manner as described in Example
36.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(isobutyryl)thio]-1-butenyl]formam
ide hydrochloride
The solution of the sodium salt of Compound B (5.2 g) in water (100 ml) was
saturated with sodium sulfate. N hydrochloric acid was added dropwise to
the solution to adjust the pH to 3, and the mixture was partitioned with
chloroform to extract fluorescent substances. The chloroform layer was
dried with anhydrous sodium sulfate and concentrated under reduced
pressure. The residue obtained (3.5 g), dimethylaminopyridine (250 mg) and
the compound prepared in the above step (a) (3.84 g) were dissolved in the
mixed solvent of dichloromethane (20 ml), tetrahydrofuran (10 ml) and
acetonitrile (10 ml). After dicyclohexylcarbodiimide (2.28 g) was added to
the solution, the mixture was stirred at room temperature for about 16
hours. Crystalline solids deposited were removed by filtration, and the
filtrate was evaporated under reduced pressure. The residue obtained was
purified by silica gel column chromatography (CHCl.sub.3
:methanol=80:1.fwdarw.40:1). The purified product (3.98 g) was dissolved
in ethanol and added with a solution of 4N hydrochloric acid in dioxane
(3.0 ml), and the mixture was concentrated under reduced pressure. The
residue obtained was dissolved in chloroform and added dropwise to ether.
Pale green fluorescent powder obtained was collected by filtration and
dried under reduced pressure to give the title compound.
Yield: 4.79 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.23 (1H, broad), 8.59 & 8.36 (each
1H, each broad), 8.20 (1H, s), 8.13 (3H, m), 8.05 (1H, broad s), 7.80 (1H,
s), 7.66 (2H, m), 7.44 (1H, broad), 4.50 (2H, broad s), 3.97 (2H, t), 3.01
(2H, q), 2.93 (6H, s), 2.56 (1H, m), 2.53 (2H, broad), 2.48 (3H, s), 2.11
(3H, s), 1.01 (6H, d).
(c)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[2-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(isobutyryl)thio]-1-butenyl]formam
ide hydrochloride
Compound C (2.7 g), the compound prepared in the above step (a) (2.8 g) and
dimethylaminopyridine (250 mg) were suspended in tetrahydrofuran (20 ml)
and dichloromethane (20 ml). Dicyclohexylcarbodiimide (1.77 g) was
dissolved in this solution, and left standing at room temperature for
about 12 hours. After white insolubles were removed by filtration, the
filtate was concentrated under reduced pressure. The residue obtained was
purified by silica gel column chromatography (developed with CHCl.sub.3
:acetone=10:1, then eluted with CHCl.sub.3 :methanol=40:1.fwdarw.35:1).
The purified product (2.78 g) was dissolved in ethanol (50 ml) and added
with 1N hydrochloric acid (4.0 ml), and the mixture was concentrated under
reduced pressure. Ethanol was added again to the residue obtained. And
then the mixture was concentrated under reduced pressure to give a
yellowish white amorphous product. The product was dissolved in chloroform
and added dropwise to ether. Greenish white powder deposited was collected
by filtration and dried under reduced pressure to give the title compound.
Yield: 2.60 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.20 (1H, broad), 8.57 (1H, d), 8.46
& 8.27 (each 1H, each d), 8.17 (1H, s), 8.09 (1H, d), 8.04 (2H, broad),
7.81 (1H, s), 7.60 (2H, m), 7.26 (1H, d), 4.49 (2H, broad), 3.88-3.80 (3H,
m), 3.76 (1H, m), 2.83 (6H, s), 2.58 (1H, m), 2.46 (3H, s), 2.37 (2H,
broad), 2.10 (3H, s), 1.11 (3H, d), 1.02 (6H, d).
Example 38
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(pivaloy
l)thio]-1-butenyl]formamide
This compound was synthesized in the same manner as described in Example
36.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(pivaloyl)thio]-1-butenyl]formamid
e hydrochloride
The solution of the sodium salt of Compound B (5.2 g) in water (100 ml) was
saturated with sodium sulfate. 4N hydrochloric acid was added dropwise to
the solution to adjust the pH to 3. The mixture was then partitioned with
chloroform to extract fluorescent substances. The chloroform layer was
dried with anhydrous sodium sulfate and concentrated under reduced
pressure. The residue obtained (3.60 g), dimethylaminopyridine (250 mg)
and the compound prepared in the above step (a) (4.03 g) were dissolved in
the mixed solvent of dichloromethane (30 ml) and tetrahydrofuran (15 ml).
Dicyclohexylcarbodiimide (2.4 g) was added to the solution, and the
mixture was stirred at room temperature for about 14 hours. After
crystalline solids deposited were removed by filtration, the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=70:1.fwdarw.40:1).
The purified product (5.08 g) was dissolved in ethanol and added with a
solution of 4N hydrochloric acid in dioxane (1.9 ml), and then the mixture
was concentrated under reduced pressure. The residue obtained was
dissolved in chloroform and added dropwise to ether. Pale green
fluorescent powder obtained was collected by filtration and dried under
reduced pressure to give the title compound.
Yield: 4.67 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.27 (1H, broad), 8.48 & 8.26 (each
1H, each d), 8.19 (1H, s), 8.10 (1H, dd), 8.06 (3H, broad m), 7.79 (1H,
s), 7.63 & 7.59 (each 1H, each t), 7.28 (1H, d), 4.50 (2H, broad), 3.96
(2H,.t), 3.01 (2H, q), 2.84 (6H, s), 2.46 (3H, s), 2.41 (2H, t), 2.12 (3H,
s), 1.06 (9H, s).
(c)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(pivaloyl)
thio]-1-butenyl]formamide hydrochloride
The compound prepared in the above step (a) (3.8 g), Compound C (3.4 g) and
dimethylaminopyridine (250 mg) were suspended in a mixed solvent of
tetrahydrofuran (20 ml) and dichloromethane (20 ml).
Dicyclohexylcarbodiimide (2.3 g) was dissolved in this solution and left
standing at room temperature for about 11 hours. After white insolubles
were removed by filtration, the filtrate was concentrated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :methanol=50:1.fwdarw.40:1). The purified
product (4.65 g) was dissolved in ethanol (100 ml) and added with 1N
hydrochloric acid (6.9 ml), and then the mixture was concentrated under
reduced pressure. Ethanol was added to the residue obtained. The mixture
was concentrated again under reduced pressure to give a yellowish white
amorphous product. The product was dissolved in chloroform and added
dropwise to ether. Greenish white powder deposited was collected by
filtration and dried under reduced pressure to give the title compound.
Yield: 4.07 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.24 (1H, broad), 8.58 (1H, d), 8.47
& 8.28 (each 1H, each d), 8.19 (1H, s), 8.09 (1H, dd), 8.06 (2H, broad),
7.81 (1H, s), 7.60 (2H, q), 7.28 (1H, d), 4.49 (2H, broad), 3.83 (2H, m),
3.75 (1H, m), 2.84 (6H, s), 2.46 (3H, s), 2.34 (2H, broad), 2.11 (3H, s),
1.10 (3H, d), 1.07 (9H, s).
Example 39
(a)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-hydroxy-1-methyl-2-[(benzoyl
)thio]-1-butenyl]formamide
This compound was synthesized in the same manner as described in Example
36.
(b)
N-[(4-amino-2-methyl-5-pyrimidyl)methyl]-N-[4-[3-[(5-dimethylaminonaphthyl
sulfonyl)amino]propionyl]oxy-1-methyl-2-[(benzoyl)thio]-1-butenyl]formamide
hydrochloride
The solution of the sodium salt of Compound B (5.0 g) in water (80 ml) was
saturated with sodium sulfate. 4N hydrochloricacid was added dropwise to
the solution to adjust the pH to 3, and the mixture was partitioned with
chloroform to extract fluorescent substances. The chloroform layer was
dried with anhydrous sodium sulfate and concentrated under reduced
pressure. The residue obtained (3.5 g), dimethylaminopyridine (250 mg) and
the compound prepared in the above step (a) (4.3 g) were dissolved in the
mixed solvent of dichloromethane (15 ml) and tetrahydrofuran (15 ml).
After dicyclohexylcarbodiimide (2.27 g) was added to the solution, and the
mixture was stirred at room temperature for about 16 hours. Crystalline
solids deposited were removed by filtration, and the filtrate was
evaporated under reduced pressure. The residue obtained was purified by
silica gel column chromatography (CHCl.sub.3 :methanol=60:1.fwdarw.40:1).
The purified product (2.82 g) was dissolved in ethanol and added with a
solution of 4N hydrochloric acid in dioxane (1.1 ml). And then the mixture
was concentrated under reduced pressure. The residue obtained was
dissolved in chloroform and added dropwise to ether. Pale green
fluorescent powder obtained was collected by filtration and dried under
reduced pressure to give the title compound.
Yield: 2.70 g.
.sup.1 H-NMR (in DMSO-d.sub.6): .delta.9.14 (1H, broad), 8.48 & 8.27 (each
1H, each d), 8.20 (1H, s), 8.10 (1H, dd), 8.06 (3H, broad m), 7.92 (1H,
s), 7.74-7.50 (7H, m), 7.28 (1H, d), 4.52 (2H, broad), 4.01 (2H, t), 3.01
(2H, q), 2.84 (6H, s), 2.66 (2H, broad), 2.40 (2H, t), 2.22 & 2.18 (each
3H, each s).
Example 40
(a) N-(2-hydroxyethyl)-N-[2-(2-propyl)dithio]ethyl]acetamide
In the solution of Intermediate 12 (8.0 g) in distilled water (50 ml) was
dissolved Sodium hydroxide (2.8 g). Sodium 2-propylthiosulfate (10 g) in
the form of powder was dissolved in the solution. The oily product
deposited was extracted with ethyl acetate and dried over anhydrous sodium
sulfate. The solvent was then evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=50:1.fwdarw.30:1) to give the title compound.
Yield: 4.2 g.
.sup.1 H-NMR (in CDCl.sub.3, 2:1 isomeric mixture): .delta.3.78 (2H, m),
3.62 (2H, m), 3.54 & 3.50 (1.33 & 0.67H, each t), 3.21 (1H, t), 3.03 (1H,
m), 2.91 & 2.81 (0.67 & 1.33H, each t), 2.18 & 2.15 (2 & 1H, each s), 1.33
(6H, d).
(b)
N-[2-[2-[(5-dimethylaminonaphthylsulfonyl)amino]ethylaminocarbonyloxy]ethy
l]-N-[2-[(2-propyl)dithio]ethyl]acetamide hydrochloride
In the solution of the compound prepared in the above step (a) (2.0 g) in
tetrahydrofuran (15 ml) was dissolved carbodiimidazole (1.54 g) in the
form of powder in a water bath at 40--C. (about 5 minutes). Compound A
(2.9 g) was added to the solution, and the mixture was stirred at room
temperature for about 15 minutes. After insolubles were removed by
filtration, the filtrate was evaporated under reduced pressure. The
residue obtained was purified by silica gel column chromatography
(CHCl.sub.3 :methanol=60:1.fwdarw.40:1). The fluorescent yellowish green
residue (3.76 g) thus obtained was dissolved in chloroform (50 ml), added
with a solution of 4N hydrochloric acid in dioxane (1.7 ml) and
concentrated under reduced pressure. The residue obtained was dissolved in
chloroform and added dropwise to ether (200 ml). The white powdery solids
obtained were collected by filtration and dried under reduced pressure to
give the title compound.
Yield: 3.64 g.
.sup.1 H-NMR (in DMSO-d.sub.6, 1:1 isomeric mixture): .delta.8.47, 8.27 &
8.09 (each 1H, each d), 7.98 (1H, broad t), 7.61 (2H, m), 7.26 (1H, d),
7.16 & 7.09 (each 0.5H, each broad t), 4.01 & 3.95 (each 1H, each
t),-3.53-3.37 (4H, m), 3.04 (1H, m), 2.97 (2H, m), 2.87-2.76 (10H, m),
2.01 & 1.96 (each 1.5H, each s), 1.22 (6H, dd).
Example 41
(a)
N-(2-hydroxyethyl)-N-[2-[2-(tert-butyloxycarbonylamino)propionylthio]vinyl
]formamide
In the solution of sodium hydroxide (2.4 g) in distilled water (60 ml) was
dissolved Intermediate 2 (6.3 g). An active ester obtained from
N-Boc-L-alanine and N-hydroxysuccinimide (6.3 g, Novabiochem. Code No.
04-12-0003) was dissolved in a mixed solvent of acetone (15 ml) and
methanol (15 ml). The solution was added to the ice-cooled intermediate
solution described above. The solution obtained by mixing these solutions
exhibited a pH of about 7. After the solution was partitioned with
chloroform, the chloroform layer obtained was dried with anhydrous sodium
sulfate. The chloroform layer was concentrated under reduced pressure and
purified by silica gel column chromatography to give the title compound.
Yield: 6.72 g
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.33 (0.75H, s),
8.11 (0.25H, s), 6.29 (0.25 H, d), 6.49 (0.75H, d), 6.21 (0.75H, d), 6.25
(0.25H, d), 4.91 (1H, broad), 4.39 (1H, m), 3.76 (4H, m), 1.47 (9H, s),
1.41 (3H, d).
Example 42
(a)
N-(2-hydroxyethyl)-N-[2-[(N-(tert-butyloxycarbonyl-2-pyrrolidyl)carbonylth
io]vinyl]formamide
In the solution of sodium hydroxide (3.2 g) in distilled water (60 ml) was
dissolved Intermediate 2 (8.4 g). An active ester obtained from
N-Boc-L-proline and N-hydroxysuccinimide (12.5 g, Novabiochem. Code No.
04-12-0076) was dissolved in a mixed solvent of acetonitrile (40 ml) and
methanol (35 ml). The solution was added to the ice-cooled above
intermediate solution in methanol (20 ml) and acetonitrile (20 ml). The
solution obtained by mixing these solutions exhibited a pH of about 7.
After the solution was partitioned with chloroform, the chloroform layer
obtained was dried with anhydrous sodium sulfate. The chloroform layer was
concentrated under reduced pressure and purified by silica gel column
chromatography (CHCl.sub.3 :methanol=80:1.fwdarw.40:1) to give the title
compound.
Yield: 8.47 g.
.sup.1 H-NMR (in CDCl.sub.3, 3:1 isomeric mixture): .delta.8.33 (0.75H, d),
8.10 (0.25H, d), 6.94 (0.25H, q), 6.48 (0.75H, t), 6.27 (0.5H, d), 6.16
(0.38H, d), 6.02 (0.12H, d), 4.46 (1H, m), 3.76 (4H, m), 3.56 (2H, m),
2.24 (1H, m), 2.06 (1H, m), 1.94 (2H, m), 1.46 (9H, d).
(b)
N-[2-[2-[(5-dimethylaminonaphthylsulfonyl)amino]propionyloxy]ethyl]-N-[2-[
(N-tert-butyloxycarbonyl-2-pyrrolidyl)carbonylthio]vinyl]formamide
In the solution of Compound C (2.6 g), the compound prepared in the above
step (a) (2.8 g) and dimethylaminopyridine (300 mg) in tetrahydrofuran (40
ml) was dissolved dicyclohexylcarbodiimide (1.8 g), and the mixture was
stirred at room temperature for about 15 hours. After white insolubles
were removed by filtration, the filtrate was concentrated under reduced
pressure. The residue obtained was purified by silica gel column
chromatography (CHCl.sub.3 :acetone=10:1.fwdarw.10:1.5).
Yield: 2.60 g.
.sup.1 H-NMR (in CDCl.sub.3, 5:1 isomeric mixture): .delta.8.55 (1H, d),
8.29 (1H, d), 8.18 (0.83H, d), 7.90 (0.17H, broad), 7.58 (1H, m), 7.52
(1H, m), 7.19 (1H, d), 6.77 (0.17H, t), 6.24 (1.7H, m), 6.18 (0.17H, d),
5.55 (1H, m), 4.45 (1H, m), 4.03-3.86 (3H, m), 3.7-3.4 (4H, m), 2.89 (6H,
s), 2.23 (1H, m), 2.03 (1H, m), 1.93 (2H, m), 1.44 (9H, d), 1.25 (3H, d).
The structures of the compounds in Examples 1-42 are shown in Tables 1 and
2. In the tables, the term "dimer" means a compound which is symmetrical
with respect to the disulfide bond.
TABLE 1
__________________________________________________________________________
Example
R1 R2 R3 R4 R5
__________________________________________________________________________
1(a)
--CH.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH.sub.2 CH.sub.3
2 --CH.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH.sub.2 CH.sub.2 CH.sub.3
3 --CH.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3).sub.2
4 --CH.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.3
5 --CH.sub.2 CH.sub.2 --OH
--H --H --H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
2
6 --CH.sub.2 --COOH
--H --H --H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
6
7 --CH.sub.2 CH.sub.2 --OH
--CH.sub.3
--H --H --SCH(CH.sub.3).sub.2
8 --CH.sub.2 CH.sub.2 --OH
--H --CH.sub.3
--H --SCH(CH.sub.3).sub.2
9 --CH.sub.2 CH.sub.2 --OH
--H --CH.sub.3
--H dimer
10 --CH.sub.2 CH.sub.2 --OH
--CH.sub.3
--H --CH.sub.3
--SCH(CH.sub.3).sub.2
11 --CH.sub.2 CH.sub.2 --OH
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3).sub.2
12 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH.sub.2 CH.sub.2 CH.sub.3
13 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH(CH.sub.3).sub.2
14 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.3
15 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --S(CH.sub.2).sub.4 CH.sub.3
16 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
.
17 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH(CH.sub.2 CH.sub.3).sub.2
18 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SC.sub.6 H.sub.11
19 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SC.sub.12 H.sub.25
20 --CH.sub.2 CH.sub.2 N(CH.sub.3).sub.2
--CH.sub.3
--CH.sub.2 CH.sub.2 OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
__________________________________________________________________________
TABLE 2
__________________________________________________________________________
Example
R1 R2 R3 R4 R5
__________________________________________________________________________
21 --CH.sub.2 CH.sub.2 NC.sub.4 H.sub.8
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
22 --CH.sub.2 CH.sub.2 NHBoc
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H dimer
23 --CH.sub.2 CH.sub.2 --OH
--H --H --H --SCH(CH.sub.3).sub.2
24 --CH.sub.2 CH.sub.2 --OH
--H --H --H --COCH(CH.sub.3).sub.2
25 --CH.sub.2 CH.sub.2 --OH
--H --H --H --COC(CH.sub.3).sub.3
26 --CH.sub.2 CH.sub.2 --OH
--H --H --H --CO--C.sub.6 H.sub.5
27 --CH.sub.2 CH(CH.sub.3)OH
--CH.sub.3
--H --H --COC(CH.sub.3).sub.3
28 --CH.sub.2 CH.sub.2 N(CH.sub.3).sub.2
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --COC(CH.sub.3).sub.3
29 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --S--CH.sub.2 --C.sub.4 H.sub.7 O
30 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3).sub.2
31 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH.sub.2 CH.sub.2 CH.sub.3
32 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
33 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --SCH(CH.sub.2 CH.sub.3).sub.2
34 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --S(CH.sub.2).sub.4 CH.sub.3
35 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --S--CH.sub.2 --CH.dbd.CH.sub.2
36 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --CO--CH.sub.2 CH.sub.2 CH.sub.3
37 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --CO--CH(CH.sub.3).sub.2
38 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --CO--C(CH.sub.3).sub.3
39 --CH.sub.2 --C.sub.5 H.sub.6 N.sub.3
--CH.sub.3
--CH.sub.2 CH.sub.2 --OH
--H --CO--C.sub.6 H.sub.5
40* --CH.sub.2 CH.sub.2 --OH
--H --H --H --SCH(CH.sub.3)CH.sub.2 CH.sub.2 CH.sub.3
41 --CH.sub.2 CH.sub.2 --OH
--H --H --H --CO--CH(CH.sub.3)--NH-Boc
42 --CH.sub.2 CH.sub.2 --OH
--H --H --H --CO--C.sub.4 H.sub.7 N-Boc
__________________________________________________________________________
*In the compound of Example 40, the bond between carbon atoms to which
R.sup.2 and R.sup.3 are bonded, respectively, is a single bond, while in
other compound the bond represents a double bond.
In vivo disposition test 1
The distribution and residence in brain of the compounds according to the
present invention were evaluated as follows.
The aqueous solutions of the compounds prepared in Examples 3 (b), 30 (b),
29 (b) and 35 (b) were respectively administered at a dose of 5 mg/kg on
the basis of Compound D of which both Boc and pivaloyl groups have been
deprotected (referred to herein after as DOPA) into the femoral vein of SD
rats. Blood samples were taken out from abdominal aorta after a certain
period of collected. Immediately after the exsanguination, whole brain was
removed. Experiments were carried out with 3 rats at each blood sampling
point.
The DOPA concentration in the whole brain was determined in accordance with
the method "New Biochemical Experiment Course, Vol. 11, Neurobiochemistry,
p. 289-290 (edited by Biochemical Society, Japan, published by Tokyo
Kagaku-Dojin, 1990)." As the controls, the Boc-eliminated Compound D and
DOPA were administered in the same manner as above to determine the DOPA
concentration.
The DOPA concentrations in brain and plasma are shown in FIG. 1 and 2,
respectively. It is understood from these figures that DOPA is transferred
into brain and stays within brain by the compound according to the present
invention.
Furthermore, the area under the concentration of DOPA in brain is shown in
the following table.
__________________________________________________________________________
Area under the curve of concentration (AUC) of DOPA in
brain (ng .multidot. min/g)
Boc-
Time
Example
Example
Example
Example elimination
(hr)
3 (a)
30 (b)
35 (b)
29 (b)
DOPA
of Compound D
__________________________________________________________________________
0-4
82476 10626
7151 6035
3277
0-2 34595 6659 6035
3169
__________________________________________________________________________
In vivo disposition test 2
The compounds of Examples 31 (b) and 32 (b) (both supporting Compound A),
the compounds of Examples 34 (b) and 38 (b) (both supporting Compound B),
and the compounds of Examples 32 (d) and 38 (c) (both supporting Compound
C) were respectively dissolved in physiological saline to prepare a 30
.mu.mol/ml (in this connection, those which were hardly soluble in only
physiological saline were dissolved in physiological saline containing
dimethylsulfoxide, propylene glycol or hydroxypropyl-.beta.-cyclodextrin).
These solutions which have passed through a 0.45 .mu.m filter were used
for the following experiments.
The solution was administered at a dose of 30 .mu.mol/kg for each compound
into the femoral vein of male SD rats. Blood samples were collected with a
heparinized syringe from abdominal aorta after a certain period of time.
Immediately after exsanguination, whole brain including cerebrum,
cerebellum and brain stem was removed from skull part. Whole blood was
immediately ice-cooled and centrifuged (3,000 rpm, 10 minutes), and then
plasma was collected and freeze-stored at -20.degree. C. The whole brain
was rinsed with physiological saline and freeze-stored at -20.degree. C.
The administration of the solution and the dissection were conducted under
anesthetization with ether.
Samples for the determination were prepared as follows.
After plasma (1 ml) was first stirred, acetonitrile (1.5 ml) and 7% (w/v)
perchloric acid (0.1 ml) were added. The mixture was stirred and left
standing for 30 minutes, and was subsequently centrifuged
(488,000.times.g, 45 minutes). The supernatant was obtained as a sample
for determination.
Brain after thawing was weighed and homogenized (20,000 rpm, 1 minute).
After a 7% (w/v) hydrochloric acid-acetonitrile solution (the weight of
the brain weighed.times.5/2 ml) was added, the mixture was homogenized
(20,000 rpm, 1 minute), stood for 30 minutes and centrifuged
(12,100.times.g, 45 minutes) to collect the supernatant. The supernatant
was further centrifuged (488,000.times.g, 45 minutes), and the supernatant
was obtained as a sample for determination.
In addition to the sample for determination obtained as above, the samples
for determination were also prepared for Whole plasma and whole brain
taken from the subject animals to which the compound according to the
present invention was not administered. The internal standard solution was
added in an amount of 1 ml and the weight of the brain weighed.times.5/2
ml, respectively.
Samples for determination obtained as above were analyzed by
high-performance liquid chromatography with a fluorescence detector to
isolate and quantitatively determine the fluorescent substance, that is
Compound A C. The concentrations of these substances were measured with
the calibration curve of the standard solution in which the internal
standard solution was added.
The results are shown in FIGS. 3-5. It is understood apparently from these
figures that Compounds A-C are delivered and retained into brain by the
compound according to the present invention.
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